Title: Enhancement of protein production via the strong DIT1 terminator and two RNA-binding proteins in Saccharomyces cerevisiae
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- Aleksi Salo
- 4 vuotta sitten
- Katselukertoja:
Transkriptio
1 Title: Enhancement of protein production via the strong DIT1 terminator and two RNA-binding proteins in Saccharomyces cerevisiae Authors: Yoichiro Ito, Takao Kitagawa, Mamoru Yamanishi, Satoshi Katahira, Shingo Izawa, Kenji Irie, Makoto Furutani-Seiki and Takashi Matsuyama Supplementary Figures S1-S8 and Tables S1-S5.
2 Yeast genomic tiling collection pool X DIT1t strain TDH3pro GFP DIT1t X Y Z gene X gene Y gene Z Supplymentary Figure S1. Scheme of screening for DIT1t-activating factors by using the Yeast Genomic Tiling Collection. The DIT1t strain, which expresses GFP under the control of the TDH3 promoter (TDH3pro) and the DIT1 terminator (DIT1t), was transformed with plasmid DNAs isolated from the yeast genomic tiling collection. The GFP fluorescence intensity of the transformants was then analyzed by flow cytometry and transformants with the highest GFP fluorescence intensity (marked with grey shading) were selected for further study. Plasmid DNAs containing multiple genes (marked as genes X, Y, Z) were then isolated from these transformants, and individual candidate genes (e.g., gene X) were cloned by gap-repair complementation, resulting in identification of the enhancing genes for the DIT1 terminator.
3 GFP/SS Log phase * * * * A43410LogA A43410LogA A43410LogA A43410LogA A43410LogA A43410LogA A43410LogB A43410LogB A43410LogB A43410LogB A43410LogB A43410LogB A43410LogC A43410LogC A43410LogC A43410LogC A43410LogC A43410LogC A43410LogD A43410LogD A43410LogD A43410LogD A43410LogD A43410LogD A43410LogE A43410LogE A43410LogE A43410LogE A43410LogE A43410LogE A43410LogF A43410LogF A43410LogF A43410LogF A43410LogF A43410LogF A43410LogG A43410LogG A43410LogG A43410LogG A43410LogG A43410LogG A43410LogH A43410LogH A43410LogH A43410LogH A43410LogH A43410LogH GFP/SS Log phase A43416LogA A43416LogA A43416LogA A43416LogA A43416LogA A43416LogA A43416LogB A43416LogB A43416LogB A43416LogB A43416LogB A43416LogB A43416LogC A43416LogC A43416LogC A43416LogC A43416LogC A43416LogC A43416LogD A43416LogD A43416LogD A43416LogD A43416LogD A43416LogD A43416LogE A43416LogE A43416LogE A43416LogE A43416LogE A43416LogE A43416LogF A43416LogF A43416LogF A43416LogF A43416LogF A43416LogF A43416LogG A43416LogG A43416LogG A43416LogG A43416LogG A43416LogG A43416LogH A43416LogH A43416LogH A43416LogH A43416LogH A43416LogH Supplementary Figure S2. Raw data used in the first screening. Candidate clones were located in the upper-row data set. No candidate clone was detected in the lower-row data set..
4 12 * * Flruorescence Intensity (a.u.) 6 * * 0 pgp564 7F9 10A10 11E10 12B3 Supplementary Figure S3. Effects of plasmids on the DIT1t strain. DNA of each plasmid vector pgp564 (Control), 7F9, 10A10, 11E10, or 12B3 (Extended Data Table 1) was transformed into the DIT1t strain. GFP fluorescence intensities of transformants were measured by flow cytometry. Three or four independent experiments were performed. Error bars are standard deviations. *, P < 0.01 by Student s t-test compared with Control.
5 A 15 B 15 Flruorescence Intensity (a.u.) 10 5 * control NAB6 overexpression Flruorescence Intensity (a.u.) 10 5 * control PAP1 overexpression 0 DIT1t RPL41Bt RPL15At RPL3t IDP1t PGK1t 0 DIT1t RPL41Bt RPL15At RPL3t IDP1t PGK1t C 15 D 15 control control Flruorescence Intensity (a.u.) 10 5 * MOS2 overexpression Flruorescence Intensity (a.u.) 10 5 * * CCC1 overexpression * * * * 0 DIT1t RPL41Bt RPL15At RPL3t IDP1t PGK1t 0 DIT1t RPL41Bt RPL15At RPL3t IDP1t PGK1t Supplementary Figure S4. Effects of enhancement of the top-five-ranked terminators. (A) Strains DIT1t, RPL41Bt, RPL15At, RPL3t, IDP1t, and PGK1t harboring GFP under the control of the TDH3 promoter and each terminator were transformed with (A) pgp564 or pgp564-nab6 vector; (B) pgp564 or pgp564-pap1 vector; (C) pgp564 or pgp564-mos2 vector; or (D) pgp564 or pgp564-ccc1 vector. The resulting GFP fluorescence intensities were measured by flow cytometry. Three or four independent experiments were performed. Error bars are standard deviations. *, P < 0.01 by Student s t-test compared with pgp564.
6 10 NAB6 Flruorescence Intensity (a.u.) 5 * nab6 0 DIT1t RPL41Bt RPL15At RPL3t IDP1t PGK1t Supplementary Figure S5. Effects of NAB6 deletion on the activities of the top-five-ranked terminators. Strains DIT1t, RPL41Bt, RPL15At, RPL3t, IDP1t, and PGK1t harboring GFP under the control of the TDH3 promoter. The resulting GFP fluorescence intensities were measured by flow cytometry. Four independent experiments were performed. Error bars are standard deviations. *, P < 0.01 by Student s t-test compared between NAB6 and nab6 background.
7 A UAAAGUAAGAGCGCUACAUU GGUCUACCUUUUUGUUCUUU UACUUAAACAUUAGUUAGUU CGUUUUCUUUUUCUCAUUUU d1 d2 UUUAUGUUUCCCCCCCAAAG UUCUGAUUUUAUAAUAUUUU AUUUCACACAATTCCAUUUA ACAGAGGGGGAATAGAUUCU d3 d4 d5 UUAGCUUAGAAAAUUAGUGA UCAAUAUAUAUUUGCCUUUC UUUUCAUC d6 B 16 Fluorescent Intensity (a.u.) 8 Control NAB6 overexpression 0 WT d1 d2 d3 d4 d5 d6 PAP1 overexpression Supplementary Figure S6. Deletion analysis of the DIT1 terminator for identification of the Pap1p- and Nab6p-recruiting region. (A) Sequence of the DIT1 3 -UTR. Deletion sites (d1 to d6) are underlined. (B) Strains harboring the DIT1 terminator region (Control) with deletion of d1 to d6 were constructed and then transformed with pgp564 (white), pgp564-nab6 (gray), or pgp564-pap1 (blue). The GFP fluorescence intensities were measured by flow cytometry. Three independents experiments were performed. Error bars are standard deviations.
8 PGK1t DIT1t-inserted PGK1t GATCA TTTTTTTCTT GATCAAAAACATTAGTTAGTTCGTTTTTTTCTT PGK1t 17 TTGAA TCGATAGATCA 29 TTTTTTTCTT DIT1t-substituted PGK1t CATTAGTTAGTTCG Supplementary Figure S7. Terminator regions of DIT1t-inserted PGK1t (A) and DIT1t-substituted PGK1t (B). The position of insertion or substitution of the sequence of DIT1t is indicated above the sequence of PGK1t. The candidate cis sequence, AGTTCG, is indicated in red.
9 2.0 Relative FI 1.0 ASP3 PIR1 FMP 52 MFA1 SCW4 TIR2 YDR 133C RPL 26B RPL 26A HSP 150 YLR 285C SUC2 GPB1 YGP1 SUI1 YDL 159W DDR2 YPR 053C Control NAB6 overexpression PAP1 overexpression NAB6+PAP1 overexpression Supplementary Figure S8. Activation of terminators of Nab6p-binding mrna genes by overexpression of NAB6 and PAP1. Strains harboring each terminator region were constructed and then transformed with pgp564 (white), pgp564-nab6 (gray), pgp564-pap1 (blue), or pgp564-nab6-pap1 (green). GFP fluorescence intensities were then measured by flow cytometry. Three independent experiments were performed. Error bars are standard deviations.
10 Supplementary Table S1. Genes included in genomic locus Plasmid name Chomosome location Genes in the genomic locus 7F9 chr k-960k 10A10, 10B2, 10D6, 10F2 chr k-450k 11E10 chr k-583k 12B3, 12B4 chr k-38k *: Asterisk shows enhanced genes for the DIT1 terminator YHB1, MOS2 *, SPG1, YGR237C, KEL2, PEX2 AUR1, MRP17, DID4, MET14, VPS1, PAP1 *, ECM9, YKR005C MSC3, CCC1 *, RSA3, UTP13, YLR222C-A YML119W, NGL3, NAB6 *, YML116W-A
11 Supplementary Table S2. DNA Sequences of terminator regions. Name d1 d2 d3 d4 d5 d6 d7 d8 d9 DNA sequence TAAAGTAAGAGCGCTACATTTTTGTTCTTTTACTTAAACATTAGTTAGTT CGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAGTTCTGATTTT ATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAATAGATTCT TTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTTTCATCTT TTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT CGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAGTTCTGATTTT ATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAATAGATTCT TTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTTTCATCTT TTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAGTTCGTTTTCTTTTTCTCATTTTCCCCCCAAAGTTCTGATTTT ATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAATAGATTCT TTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTTTCATCTT TTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATTTCACACAATTCCATTTAACAGAGGGGGAATAGATTCT TTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTTTCATCTT TTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAAATAGATTCT TTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTTTCATCTT TTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGATCAATATATATTTGCCTTTCTTTTCATCTT TTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TAAAGTAAGAGCGCTACATTGGTCTACCTTTTTCTTTTACTTAAACATTA GTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAGTTC TGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAAT AGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTT TCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TAAAGTAAGAGCGCTACATTGGTCTACCTTTTTGTTTTACTTAAACATTA GTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAGTTC TGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAAT AGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTT TCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TAAAGTAAGAGCGCTACATTGGTCTACCTTTTTGTTCTACTTAAACATTA GTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAGTTC TGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAAT AGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTT TCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT
12 d10 TAAAGTAAGAGCGCTACATTGGTCTACCTTTTTGTTCTTTTTAAACATTA GTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAGTTC TGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAAT AGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTT TCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT d11 TAAAGTAAGAGCGCTACATTGGTCTACCTTTTTGTTCTTTTACTACATTA GTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAGTTC TGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAAT AGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTT TCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT d12 TAAAGTAAGAGCGCTACATTGGTCTACCTTTTTGTTCTTTTACTTAATTA GTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAGTTC TGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAAT AGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTT TCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT d13 GTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAGTTC TGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAAT AGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTT TCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT d14 TTAAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAGTTC TGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAAT AGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTT TCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT d15 TTAGTTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAGTTC TGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAAT AGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTT TCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT d16 TTAGTTAGTTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAGTTC TGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAAT AGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTT TCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT d17 TTAGTTAGTTCGTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAGTTC TGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAAT AGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTT TCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT d18 TTAGTTAGTTCGTTTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAGTTC TGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAAT AGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTT TCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT d19 TTAGTTAGTTCGTTTTCTTCTCATTTTTTTATGTTTCCCCCCCAAAGTTC TGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAAT AGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTT TCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT
13 d20 d21 TTAGTTAGTTCGTTTTCTTTTCATTTTTTTATGTTTCCCCCCCAAAGTTC TGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAAT AGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTT TCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAGTTCGTTTTCTTTTTCTTTTTTTATGTTTCCCCCCCAAAGTTC TGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGGAAT AGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTCTTT TCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT
14 Name m1 m2 m3 m4 m5 m6 m7 m8 m9 m10 m11 DNA sequence TTAGTTgGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTcGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTtGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAaTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAcTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAtTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAGaTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAGgTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAGcTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAGTaCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAGTgCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT Position of the mutated nucleotide The 5th A to G in "AGTTAGTTCG" The 5th A to C in "AGTTAGTTCG" The 5th A to T in "AGTTAGTTCG" The 6th G to A in "AGTTAGTTCG" The 6th G to C in "AGTTAGTTCG" The 6th G to T in "AGTTAGTTAG" The 7th T to A in "AGTTAGTTAG" The 7th T to G in "AGTTAGTTAG" The 7th T to C in "AGTTAGTTCG" The 8th T to A in "AGTTAGTTAG" The 8th T to G in "AGTTAGTTAG"
15 m12 m13 m14 m15 m16 m17 m18 m19 m20 m21 m22 TTAGTTAGTcCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAGTTaGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAGTTgGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAGTTtGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAGTTCaTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAGTTCcTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTTAGTTCtTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTgGTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTcGTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTtGTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAaTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT The 8th T to C in "AGTTAGTTAG" The 9th C to A in "AGTTAGTTCG" The 9th C to G in "AGTTAGTTCG" The 9th C to T in "AGTTAGTTCG" The 10th G to A in "AGTTAGTTCG" The 10th G to C in "AGTTAGTTCG" The 10th G to T in "AGTTAGTTCG" The 1st A to G in "AGTTAGTTCG" The 1st A to C in "AGTTAGTTCG" The 1st A to T in "AGTTAGTTCG" The 2nd G to A in "AGTTAGTTCG"
16 m23 m24 m25 m26 m27 m28 m29 m30 TTAcTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAtTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGaTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGgTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGcTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTaAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTgAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT TTAGTcAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT The 2nd G to C in "AGTTAGTTCG" The 2nd G to T in "AGTTAGTTCG" The 3rd T to A in "AGTTAGTTCG" The 3rd T to G in "AGTTAGTTCG" The 3rd T to C in "AGTTAGTTCG" The 4th T to A in "AGTTAGTTCG" The 4th T to G in "AGTTAGTTCG" The 4th T to C in "AGTTAGTTCG"
17 Name DNA sequence length DIT1t TTAGTTAGTTCGTTTTCTTTTTCTCATTTTTTTATGTTTCCCCCCCAAAG TTCTGATTTTATAATATTTTATTTCACACAATTCCATTTAACAGAGGGGG AATAGATTCTTTAGCTTAGAAAATTAGTGATCAATATATATTTGCCTTTC TTTTCATCTTTTCAGTGATATTAATGGTTTCGAGACACTGCAATGGCCCT 250 bp PGK1t DIT1t-inserted PGK1t DIT1tsubstituted PGK1t ATTGAATTGAATTGAAATCGATAGATCAATTTTTTTCTTTTCTCTTTCCC CATCCTTTACGCTAAAATAATAGTTTATTTTATTTTTTGAATATTTTTTA TTTATATACGTATATATAGACTATTATTTATCTTTTAATGATTATTAAGA TTTTTATTAAAAAAAAATTCGCTCCTCTTTTAATGCCTTTATGCAGTTTT TTTTTCCCATTCGATATTTCTATGTTCGGGTTCAGCGTATTTTAAGT ATTGAATTGAATTGAAATCGATAGATCAAAAACATTAGTTAGTTCGTTTT TTTCTTTTCTCTTTCCCCATCCTTTACGCTAAAATAATAGTTTATTTTAT TTTTTGAATATTTTTTATTTATATACGTATATATAGACTATTATTTATCT TTTAATGATTATTAAGATTTTTATTAAAAAAAAATTCGCTCCTCTTTTAA TGCCTTTATGCAGTTTTTTTTTCCCATTCGATATTTCTATGTTCGGGTTC AGCGTATTTTAAGT ATTGAATTGAATTGAAACATTAGTTAGTTCGTTTTTTTCTTTTCTCTTTC CCCATCCTTTACGCTAAAATAATAGTTTATTTTATTTTTTGAATATTTTT TATTTATATACGTATATATAGACTATTATTTATCTTTTAATGATTATTAA GATTTTTATTAAAAAAAAATTCGCTCCTCTTTTAATGCCTTTATGCAGTT TTTTTTTCCCATTCGATATTTCTATGTTCGGGTTCAGCGTATTTTAAGT 247 bp 264 bp 249 bp
18 Supplementary Table S3. List of genes, pimers and amplified terminator regions of NAB6 -binding mrna genes. Systematic Name Gene Name 5'-primer 3'-primer Sequence of amplified region 1 YLR155C ASP3-1 AGCCTGATCAGTGACTTTTCATC GCGGCCATATCTACCAGAAA AGCCTGATCAGTGACTTTTCATCACACTGTGTTTTTATATAGTTCTTAGTAGTAAATATACTCTATAATTTCTACATCCTTTTC ATGATGTGCCAGTTAAGCTATTTTTAGCAAACAAAAAGATGAATTTGAGTATTGAGCACATCATTCACTGTTCAGCTGATACAA AACATATGATTTCTAATATTCACATAGGAAAAATCGCTCCAAACATCGAAGTATCATCGCTACTTTGGTCTTTTATCTGCTTCT CTTCTCTTCGCTCTTTTCACTGCTCGTGCCGCTGCAACGCTCCAACTCAGTTCTATCTCAGCTCCTTCCCCTTATCCCTTGAGC AGTGTCAAGGCGAGGCTGCGTGATGTATTCGTGCGCGAAAAAAAAAACAACTGCAGCACCTGAGTTTCGCGTATGGTCACCCAC TACACTACTCGGTCAGGCTCTTACCAGCTTAACTACAGTTGATCGGACGGGAAACGGTGCTTTCTGGTAGATATGGCCGC 2 YKL164C PIR1 TCATAAGTTTACCTCTCATAGTCGCA GGCTCCACGGTACACAAAGT TCATAAGTTTACCTCTCATAGTCGCATGAAAACATAAAGTCATCGTAGCATATTCTGTACATTAGTCATTTCTGTCTCCAAAGG GCATAATTTGGGTCGGTTCATTTTTTTCTTCTTAATTCGTCTCTTCATTTCATAAAGCTTTCTTTTGAAACCTATGAAATGAGC TTTACAGTTCTTCACTCTTTTTTTTAAAGTTTGTTTATTTTTTTATACTATATACATTATTATTTCTAAAAAATCATTATAAAT TACGTTAAAAGCGCACCTTATACTTTGATAGAGTTTTCCCAGAAAAAAAACCAACGAATTTGATAGTACCAAGACTTCACGGGA TTGAAATCGGATGCAACTTCAACTATTCCTGGGACAGAATATTTCTCAGACATTACTTTGTGTACCGTGGAGCC 3 YER004W FMP52 ATCAACCAACGGCTTCTTCT TGTGAGTCCTGGTTTGTTGC ATCAACCAACGGCTTCTTCTCGCAGCTACAGTTGTATTTTACTGTGACGTACTTTTGCGCGCGTACGGGTTCAATGAATTACGG ATTTTCAACAACTTCACACAATCATAGCTTTTTTTACTTTTTCGCCTCATTATTTTCTTTAAAGAAACGATGATAAACAGAGAT TTATCGTCATGTTATTTCGTTCGTATATATCGTATTTGCGTTTTAGCTGTATAAAGTAAAATACTATATCTAACGTGGCTTTTT TTATTTCCCCGTCTGCCCTCTTATGCTCATAGAAAACACTAATGATCGGTTTGGTATCGTCATAGATGCAGGGTCTTCGGGTTC GAGAATCCATGTGTTTAAGTGGCAGGACACGGAATCGCTTCTTCATGCAACAAACCAGGACTCACA 4 YDR461W MFA1 GTTATTGCTTAGTTTCTGCGTACA TAGATCCGCCCTCTTACGAA 5 YGR279C SCW4 GTCTACTCAATTGTAAAAAAAAACAAA CCAAGAGGATCGAGAAACCA GTTATTGCTTAGTTTCTGCGTACAAAAACGTTGTTCTCCCTCCTTTATCTTCCTTTTCCGCTACACCAATATATCATGTTTGTT CGTAATATTTCTTTTTAGACCTAATAATAAATATCCTAAGTAACTATATTATATAAAATATTTTGATACCCTGTACCTGCTTTT GTTATCGTTGTACATCCATGCACACGCTCATATATATATATATATATATATATATGTATATGTACATATACCGCTTACCACCTA CCGTGAAGTATATGTAAGGGTCTTCGCACCCGGATATCCCTTGTGGGATCTTGGATGCGGATGGTGAGTGTAAACAGGCCTCAT AAAGCTGTCTCTGTGTGCGGAAGATGTCGTTTCATCGGCTTCGACCTCCTCCTTATCTTGCTTCATTGTTTCTTCGTAAGAGGG CGGATCTA GTCTACTCAATTGTAAAAAAAAACAAAAAAAATCTTGATTTAAAAAACGATACCCAATATTTTTAACTTTAAATTAAGGCTTTG ATAATCTTCTTTTTCTCTGTTATATCAAAGGACTGCTTTCTGGTTTGCCTTCGCTAATCTTTTTTTTTTTTTTTCATTTTCCTA TTTTAGTTCGTTTATGATCTCATTTGTATTTCTGTATATTTGTCTTAATACGAAATACGAGTTTTTATTTTATATTCTACCTTT CTTTTTTTTTTGGCTAAAAGCGTGTACGGTTCTATGTAAATATGTACGATTACATGTGTAATGTTTGTTTGGCCGGGCTCTTCA TCTATGCCGTTGCCTTCTTGCAGGCGGAGTCCTGGATCTGTTACGATGGTTTCTCGATCCTCTTGG 6 YOR010C TIR2 TTTTAAATGGAATTTTTAATGTTCG GCCGCCATGCTACTATGATT TTTTAAATGGAATTTTTAATGTTCGTATATACCAATTTATCTATTGAATAGTTTTTTTTAAATTTTACTGTTTCTTTTAAACCT CGAATTATTGTGAGAACTGATTTCATCAACTGGAGGCTGTAGTGTACGTTTTGAAAACGGAAATGATTTACCGTTTTCTTTTAT GATGATGCTAGGATAGGCACCTTCAAACATGCATGTCTTCCTGCCCACATTCTGAAAATATTTAACGATGGATGAAAGAAATCA ACAATAGAAAAGGCCAGTTGTCAGTTTTTATATAATACAGAGAAGTACATATGACAGTTTAAAAGTTAAGCATTAACTGGTAAG TTGAAATAATAATATAATCATAAGCGGAACGAACATTTTCGACACGTACTAAAATCATAGTAGCATGGCGGC 7 YDR133C YDR133C TTTCTTATTCTAATTCTAGG CGAGAGGAGAATCCATTAG 8 YGR034W RPL26B TTGGAATAAACTAGCTCGACTTTAAC TGTGGCCAATACTGCTTCAG 9 YLR344W RPL26A ATTATGTAATTCCCTGAAAACTTCC TTTACCCACCTTTTGACGGT 10 YJL159W HSP150 TAGTTCTTTTATCCTGATGACTTTTTC AATAGACGTAGCCACATGCAG TTTCTTATTCTAATTCTAGGAGGCAACACTCCATAACTAAATGTTTATTATAATAATATTAATATAAAGTTAATATCTCTTAGG TTACACGCAACTAAACTTTTTTTATATATACTGGTTGCATTGTATTTTTATCATATATCTTAATATAGGTTGTTTAAGGTTCGT TAATTCAAAAATAATGTTTTTTTGTTTGCCCAGTCGGTTTACGCTAATCAAGAACATGGATTCAATATCTTACATAATAAGCGG ACGATGAGTCTATCTCGGTTTCCTCTTTTCTGAAATATAATAGTTTTATATTAGTGATTCATTATTACGATCTGGCCGGCTTAT CATTGCTTCTTTTACAAACGTATCCTTTTAAGATTTTATCCGGACCACTGAGCTGGCATTTTTCTTAGCTTGCTTTTGTCTTTG TTGCACGGGCGACTGAACTTCTTGGAAAATCATGTAAGCCGCAATTGCTTACTAAGTCAAAATTCTAATGGATTCTCCTCTCG TTGGAATAAACTAGCTCGACTTTAACATAGATCATACAAAAAATACAATAAATAAATCCTGTGTTTCTTTTATGTATATTTTTT ATCAATTTCAATTTAGTTTAGTTTAGTTTCTTTAAAAATAAAATTGTAATTTCTCACATTCTGCCTTCGGGTCTCAAGGGCTGG TTGGTAGACGAATGCGCTTGTTTCTAGCTGAGCGGCAGATGCATGAATTCAATGCAAGAAAATGAGAAAACTTCCAATATTAAA ATGTGATTGTTAATTATTAGTATATTTATGACAATATATATATGTACATAGGGTGATTATGAAGATGAGTAAGATGCGAATGGA TATGAGGATATGAATCTTTTTAGAGACAATGAAGATGCAAAGAAGTAGTTAAGCACATTGACCTCAGGGTTGCTATATGCTACT TTTAGTCCGTTGGATTTCTGAAGCAGTATTGGCCACA ATTATGTAATTCCCTGAAAACTTCCTCGAAATTTATTGCACTTTTTTTACTCAAACCATTTAATAATCATATGTAAAACGTATA AAATAGTATCCATTCCACTGTACTCAGTGTCTTACGTAAAGGGCTGTATACTGGCACTACCTTTTACTATGGATATGTGCTAGC TTTTTCGGAGAAGAAATGTTCGCCTTACGCTAGAAACCGGGGTTAAGGGGGCGTGAGAATGAATATTGGCAGACAAGAACCGAG ATAGAATCTTAACACAAAAACAGTTCATCCTACTTCTACGCCTTTATACAGTTATACTGAAAGATCTTTGCATAGGGGCATAGT TTAAAATAAAATATAGGGTTGTCATAGCAAACATGCCAAATGTGCTTTCTGATGATGAAGAGCTCCTTAATGGACTAGGAAGTG AGATTATGAAACCCTCTAGACAAGGTAACCATATGGCAAGGACCGTCAAAAGGTGGGTAAA TAGTTCTTTTATCCTGATGACTTTTTCTCATTTGCATTGATTAGAAAGGAAAAAAAGAAGTGTCCTTTTCTACTACTACTCTAG TCGCATCCATTCCTTTGCATTTATCTTTTCTGCGGTTGGCCAATCCATTCTTCCGAGAATTTGGCTAGCCATACTTGATGTTTT CCCATTATTGGTTCGTTTGGCAATGCTAATTTTCTTAATTGCCCCTTATATACTCTTCCATAAAATGTTTTTTTTATAACTAAT TTTCTGTATATCATTATCTAATAATCTTATAAAATGTTAAAAAGACTTGGAAAGCAACGAGTGATCGTGACCACATAATTGCCT CGCTACACGGCAAAAATAAGCCAGTCCTAATGTGTATATTAAAGGCTGCATGTGGCTACGTCTATT 11 YLR285C-A YLR285C-A TGAGAATGCCAATTCCGC TGGTTGTTTCAACTGATTATCCA 12 YIL162W SUC2 AGGTTATAAAACTTATTGTCTTTTTT GCTTGGTATCCATTTCCCTC 13 YOR371C GPB1 TAATTAATTCCGCGGGGC TGAAATGTTGCTGTTAAACCCTT 14 YNL160W YGP1 AGATATTAAATCTAGCGAAGCATAGAG GACACAAAAGAAGGGGGTGA 15 YNL244C SUI1 GTTCAAGGCTTACGCCGAGT CATTCTTGAGAGTCTCCGGC 16 YDL159W-A YDL159W-A TCACTGTCCTCGTTATAAGTGATTC AAAACAGGTTCATTTTGCGG 17 YOL052C-A DDR2 GAAAAACGCCGCTTACTGC CTTTGGTTTCATGGCGAAAT TGAGAATGCCAATTCCGCCCACTTACTACTTTTCATGTGTTAAATAGTACGCATAAACACGAGATCTGACTTCGATTGTTTCCT CGTCTTCGTCAAACATCGGTTTCCAATTAACCATTTCAATTAGCTGAGGAACCAAATGGAACTCGTTCTTAGCCAATTCGAAGA ATTCTAAATCCTTCTCCAACAATTTTGGTCTATGCGGTGAAAATACTACCAACGCCTGACCTTTCTCAGCTAATAAATCCTTTG TTGTTTGAAGTAATTTGTGATGTTCCGTATGATTGAAGACTAAATCACTTAAAATGATTAAGTCAAACTTTCCATTATTGTTAC CTATTTTTTCGATATGTGCCAACAATGGAGAATAATCGTTTCCCCAAATATAACCCTCCGTACTGACATTATTAAAATCTTCAG GAACGTTAGACTTTATATTATAATCGATGTTCTGCATTAAATCAGGATCTGGATAATCAGTTGAAACAACCA AGGTTATAAAACTTATTGTCTTTTTTATTTTTTTCAAAAGCCATTCTAAAGGGCTTTAGCTAACGAGTGACGAATGTAAAACTT TATGATTTCAAAGAATACCTCCAAACCATTGAAAATGTATTTTTATTTTTATTTTCTCCCGACCCCAGTTACCTGGAATTTGTT CTTTATGTACTTTATATAAGTATAATTCTCTTAAAAATTTTTACTACTTTGCAATAGACATCATTTTTTCACGTAATAAACCCA CAATCGTAATGTAGTTGCCTTACACTACTAGGATGGACCTTTTTGCCTTTATCTGTTTTGTTACTGACACAATGAAACCGGGTA AAGTATTAGTTATGTGAAAATTTAAAAGCATTAAGTAGAAGTATACCATATTGTAAAAAAAAAAAGCGTTGTCTTCTACGTAAA AGTGTTCTCAAAAAGAAGTAGTGAGGGAAATGGATACCAAGC TAATTAATTCCGCGGGGCGGGGTTCGGATAATCCTTATGTGACAAGAGTGACTAAAGGAAAACGAGAAAATTTTTTCGTGGAAC TTGAAAAAAAAAAAAAAAAAAAAAAAGAAAACAGAACAAGTGAAGAAGATAAAAAAAGGTTCTATATCGCACTATACAACAATA GTAGCGTTTCCCGTGTTCTTTTCCCGTCATTAAATATATTCAAAAGTTGTTGCACATTTGATTTTACAGAAGTAATATACACTG ATTGTGCGTACGGTTAATTCGCAACTAAATTCAATCAGTACAAGAAAAATTTTTATTCCGTTCCATCATTATTGAAAGATAAAC TATCACATTGCGATAATAAACTAAAAGACACTAGGATACTGCCTTCGGATATAAACATTTAAGTTTAATTTATTACTGTTTTTT TAAGGGTTTAACAGCAACATTTCA AGATATTAAATCTAGCGAAGCATAGAGATTCTTTTTTTACCAGTCATCATAACCATTATTTTACAAAATTTCCACGCAACAGCT TTTATTTTCGTTCGTAATAGGAACCATACTTCCCATTAAAGCAGTATGATTGTTTTATACACCTTTTTATTGTTATTTAAGACA TACCCCAAATAATTAATTAATTAATTGATGAAAGAATCATATAAATTCTTGGCCATACATATTTTTTTATTATCTTTTGCTACA TTGACCATAGGTGAATATCTTCCCGGGGTGACACTCCACTGAAGTGGGAAAAAAAGAAAAGTTTTAAAACATTGCTCACCTTAA GTCTCGAAGGTCTAGCATTCACTACCTGTAAGTGTCAAGACCCCACATGTATACATGGATACAATAAAGAAACAATGTCCGTAT AATTTGTATACATTTTACTACAGATAAGGGATCTCACCCCCTTCTTTTGTGTC GTTCAAGGCTTACGCCGAGTCGCTTACTCGCTGCTCGCTTTCTTTCTTAGTGTTGTTATCCTATATAAAACGTAACTGTTTAGT TATTTAAGTCCAAGTGCATATTTACACTATAATATATAATATAACATCTCATCTTTTTTTATTTCTTTTTCTTCTTCTCCCGCC GCGGGCTTACATACAGAGATATAAGAACAAGTGGTACGTAGCATGCAGCTTCTCAAAAGAGATGAGCGTTCAGTTTTGAGTACA GGGAAGAAAGGCCAAATCGAGAGTACATTATCATGGGGTCGGGCGATTTGAATCTGTTGAAATCATGGAATCCCAAGCTTATGA AAAATAGAAAAAAAGTATGGGAGACCGAACAGGATCTAATTACCGAACAGCAGAAGCTTAACACGCGATTGAAAGAAATTGAAA AAGAACGAGAACTAAATGAACTGCTGAACGAATCGAGCAAGGATAAGCCGGAGACTCTCAAGAATG TCACTGTCCTCGTTATAAGTGATTCGTGGGGTAACGGCGATATCATACTATTATTGGCCCATCTGATGGATTAGAAGACCTACA GGTTAATGTGAAGCGACACTAATGATAAGAAAACTGCATCGGATCATATCTGTTTTTGCAGCGTGGTATATTGGTTGTTGGTCA TGTTTCAACGAAAGACTTTACAGAGAAGGAACTTGAAAGGGCTCAATCTTAACCTGCACCCAGATGTGGGCAATAATGGCCAAT TGCAGGAAAAGACAGAGACTCACCAGGGACAATCTCGAATAGAAGGCCACGTGATGTCTAACATTAATGCAATACAGAATAATA GCAACCTGTTTTTGCGAAGAGGCATAAAAAAAAAACTGACGTTGGATGCGTTTGGTGATGACCAAGCTATATCGAAACCAAACA CTGTGGTAATACAGCAACCGCAAAATGAACCTGTTTT GAAAAACGCCGCTTACTGCCACGATGATGATACCCTATTGACGTTTCTGAAATGTATAATTTCTTTTCTCATCTTCCCCTTTGA TATTCCATCTATAGGCCCCAGAGTAGTAAATTTGGTGCTTTAATTTTTTTTCTCTTCTTCCAGTTCGTCTCTATTTTTTCATTC CATTATATTTATTTATCAGTTTTACTTTCTCAAATATTCTTATATAACACTATTTCATCTACGTAACCGAAAATAAGTAGTTTA ATAACGATAATAATCATAATTGTGATAAAAATATTTACATTTTTCTTACTTGTCGCCCGCTTCGTTTTTGTATAGAACGTACTG TATTAGTTGTCCGTTCTTTTCACGTCTGAAGAGGTCGATTTCTTGCAATTTAACATCAACGTGGTATTTTCCATTATATACAAT CAAAGAATCATCGTCCAGTTCGATTTCGCCATGAAACCAAAG
19 18 YPR053C YPR053C CGCCAGAGGAAAAGCAGCC AACATTGCGATGAGATCATG CGCCAGAGGAAAAGCAGCCTTACGAGGCCAAGGCCCAGGCCGATAAGAAGAGATATGAATCCGAAAAGGAGTTATATAACGCCA CTTTGGCTTAGGGATACTTCCTCCCCGCGTTTTTGTCTGTGTGATTTAATTTCTTTTCCTCTTGTTCTTTTTGAAACTTGCTTT ATTATCTCTCTCCTTTCTATTCCTCTTTTCTCTACTGCCCTTTTTCTATATCTATACATATGATTAAATAAATAAATAAAAAAA TTGTATTATTAATATATATTTTATCATCATCGAGTGTTTTTCCTTTTATGTGGCCAGCCGGCCATGCATTAGGAACGTAGACCG AGTGCAGCTCTTTTCAGTCAACGGCAATATCAGTTTAAAGGCGTCCCCTTCATTCAAGTAGTAGCGGAACATCCGTTTCATTCT GATGAAACCCATTCCTTCATATAAGTTTAGAGCCGCCGAATTTTCCACCTCTGTTTCTAACATGATCTCATCGCAATGTT
20 Supplementary Table S4. Raw data sets of Figures 1 & 2. Population Genetic Background Overexpression Average log10 value (a.u.) of fluorecent intensity (FI) from 3000 cells full width at half maximum Average value of FI Mean value of Standard Deviatoin of NAB NAB NAB NAB NAB NAB6 NAB NAB6 NAB NAB6 NAB NAB6 NAB NAB6 NAB NAB6 PAP NAB6 PAP NAB6 PAP NAB6 PAP NAB6 PAP NAB6 NAB6+PAP NAB6 NAB6+PAP NAB6 NAB6+PAP NAB6 NAB6+PAP NAB6 NAB6+PAP nab nab nab nab nab nab6 PAP nab6 PAP nab6 PAP nab6 PAP nab6 PAP t-test: Paired Two Sample for Means (p<0.01) Population 1 2 significant Mean Variance Observations 5 5 Pearson Correlation df 4 t Stat P(T<=t) one-tail E-07 t Critical one-tail P(T<=t) two-tail E-06 t Critical two-tail Population 1 3 significant Mean Variance Observations 5 5 Pearson Correlation df 4 t Stat P(T<=t) one-tail E-06 t Critical one-tail P(T<=t) two-tail E-05 t Critical two-tail
21 Population 1 4 significant Mean Variance Observations 5 5 Pearson Correlation df 4 t Stat P(T<=t) one-tail E-05 t Critical one-tail P(T<=t) two-tail t Critical two-tail Population 1 5 significant Mean Variance Observations 5 5 Pearson Correlation df 4 t Stat P(T<=t) one-tail E-07 t Critical one-tail P(T<=t) two-tail E-07 t Critical two-tail significant Mean Variance Observations 5 5 Pearson Correlation df 4 t Stat P(T<=t) one-tail E-06 t Critical one-tail P(T<=t) two-tail E-06 t Critical two-tail
22 Population Ct value of GFP primer set Ct value of TUB primer set Genetic Overexpression (Sample) (Standard) Background repetition 1 repetition 2 repetition 1 repetition 2 NAB NAB NAB NAB6 NAB NAB6 NAB NAB6 NAB NAB6 PAP NAB6 PAP NAB6 PAP NAB6 NAB6+PAP NAB6 NAB6+PAP NAB6 NAB6+PAP nab nab nab nab6 PAP nab6 PAP nab6 PAP Relative GFP mrna level to TUB1 = exp(-cttub ln2)/exp(-ctgfp ln2) Population Genetic Mean of relative SD of relative Overexpression Background mrna level mrna level 1 NAB NAB6 NAB NAB6 PAP NAB6 NAB6+PAP nab nab6 PAP t-test (P<0.01) Population 1 vs 2 significant Pooled variance Test statistic P value (two-sided) E-06 Population 1 vs 3 no difference Pooled variance Test statistic 0.9 P value (two-sided) Population 1 vs 4 significant Pooled variance Test statistic 11.7 P value (two-sided) E-07 Population 1 vs 5 no difference Pooled variance Test statistic P value (two-sided) Population 1 vs 6 no difference Pooled variance Test statistic 2.7 P value (two-sided)
23 Population Genetic Background Overexpression Average log10 value (a.u.) of fluorecent intensity (FI) from 3000 cells Average value of FI Mean value of Standard Deviatoin of Relative FI (DIT1t-- Mean value of SD of relative Mean value as a standard) relative FI FI DIT1t DIT1t DIT1t DIT1t DIT1t NAB DIT1t NAB DIT1t NAB DIT1t NAB DIT1t PAP DIT1t PAP DIT1t PAP DIT1t PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d2 NAB DIT1t-d2 NAB DIT1t-d2 NAB DIT1t-d2 NAB DIT1t-d2 PAP DIT1t-d2 PAP DIT1t-d2 PAP DIT1t-d2 PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d7 NAB DIT1t-d7 NAB DIT1t-d7 NAB DIT1t-d7 NAB DIT1t-d7 PAP DIT1t-d7 PAP DIT1t-d7 PAP DIT1t-d7 PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d8 NAB DIT1t-d8 NAB DIT1t-d8 NAB DIT1t-d8 NAB DIT1t-d8 PAP DIT1t-d8 PAP DIT1t-d8 PAP DIT1t-d8 PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d9 NAB DIT1t-d9 NAB DIT1t-d9 NAB DIT1t-d9 NAB DIT1t-d9 PAP DIT1t-d9 PAP DIT1t-d9 PAP DIT1t-d9 PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d10 NAB DIT1t-d10 NAB DIT1t-d10 NAB DIT1t-d10 NAB DIT1t-d10 PAP DIT1t-d10 PAP DIT1t-d10 PAP DIT1t-d10 PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d11 NAB DIT1t-d11 NAB DIT1t-d11 NAB DIT1t-d11 NAB DIT1t-d11 PAP DIT1t-d11 PAP
24 DIT1t-d11 PAP DIT1t-d11 PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d12 NAB DIT1t-d12 NAB DIT1t-d12 NAB DIT1t-d12 NAB DIT1t-d12 PAP DIT1t-d12 PAP DIT1t-d12 PAP DIT1t-d12 PAP significant Mean Variance E-05 Pearson Correlation Hypothesized Mean Differe 0 df 3 t Stat P(T<=t) one-tail E-05 t Critical one-tail P(T<=t) two-tail E-05 t Critical two-tail significant Mean Variance Pearson Correlation Hypothesized Mean Differe 0 df 3 t Stat P(T<=t) one-tail E-06 t Critical one-tail P(T<=t) two-tail E-06 t Critical two-tail significant Mean Variance Pearson Correlation Hypothesized Mean Differe 0 df 3 t Stat P(T<=t) one-tail E-05 t Critical one-tail P(T<=t) two-tail E-05 t Critical two-tail
25 Population Genetic Background Overexpression Average log10 value (a.u.) of fluorecent intensity (FI) from 3000 cells Average value of FI Mean value of Standard Deviatoin of Relative FI (DIT1t-- Mean value of SD of relative Mean value as a standard) relative FI FI DIT1t DIT1t DIT1t DIT1t DIT1t NAB DIT1t NAB DIT1t NAB DIT1t NAB DIT1t PAP DIT1t PAP DIT1t PAP DIT1t PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d2 NAB DIT1t-d2 NAB DIT1t-d2 NAB DIT1t-d2 NAB DIT1t-d2 PAP DIT1t-d2 PAP DIT1t-d2 PAP DIT1t-d2 PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d13 NAB DIT1t-d13 NAB DIT1t-d13 NAB DIT1t-d13 NAB DIT1t-d13 PAP DIT1t-d13 PAP DIT1t-d13 PAP DIT1t-d13 PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d14 NAB DIT1t-d14 NAB DIT1t-d14 NAB DIT1t-d14 NAB DIT1t-d14 PAP DIT1t-d14 PAP DIT1t-d14 PAP DIT1t-d14 PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d15 NAB DIT1t-d15 NAB DIT1t-d15 NAB DIT1t-d15 NAB DIT1t-d15 PAP DIT1t-d15 PAP DIT1t-d15 PAP DIT1t-d15 PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d16 NAB DIT1t-d16 NAB DIT1t-d16 NAB DIT1t-d16 NAB DIT1t-d16 PAP DIT1t-d16 PAP DIT1t-d16 PAP DIT1t-d16 PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d17 NAB DIT1t-d17 NAB DIT1t-d17 NAB DIT1t-d17 NAB DIT1t-d17 PAP DIT1t-d17 PAP
26 21 DIT1t-d17 PAP DIT1t-d17 PAP significant Mean Variance Pearson Correlation Hypothesized Mean Differe 0 df 3 t Stat P(T<=t) one-tail E-05 t Critical one-tail P(T<=t) two-tail E-05 t Critical two-tail significant Mean Variance Pearson Correlation Hypothesized Mean Differe 0 df 3 t Stat P(T<=t) one-tail E-05 t Critical one-tail P(T<=t) two-tail E-05 t Critical two-tail significant Mean Variance Pearson Correlation Hypothesized Mean Differe 0 df 3 t Stat P(T<=t) one-tail E-05 t Critical one-tail P(T<=t) two-tail E-05 t Critical two-tail significant Mean Variance Pearson Correlation Hypothesized Mean Differe 0 df 3 t Stat P(T<=t) one-tail E-05 t Critical one-tail P(T<=t) two-tail t Critical two-tail significant Mean Variance Pearson Correlation Hypothesized Mean Differe 0 df 3 t Stat P(T<=t) one-tail t Critical one-tail P(T<=t) two-tail t Critical two-tail significant Mean Variance Pearson Correlation Hypothesized Mean Differe 0 df 3 t Stat P(T<=t) one-tail E-05 t Critical one-tail P(T<=t) two-tail E-05 t Critical two-tail
27 13 16 no difference Mean Variance Pearson Correlation Hypothesized Mean Differe 0 df 3 t Stat P(T<=t) one-tail t Critical one-tail P(T<=t) two-tail t Critical two-tail no difference Mean Variance Pearson Correlation Hypothesized Mean Differe 0 df 3 t Stat P(T<=t) one-tail t Critical one-tail P(T<=t) two-tail t Critical two-tail no difference Mean Variance Pearson Correlation Hypothesized Mean Differe 0 df 3 t Stat P(T<=t) one-tail t Critical one-tail P(T<=t) two-tail t Critical two-tail
28 Population Genetic Background Overexpression Average log10 value (a.u.) of fluorecent intensity (FI) from 3000 cells Average value of FI Mean value of Standard Deviatoin of Relative FI (DIT1t-- Mean value of SD of relative Mean value as a standard) relative FI FI DIT1t DIT1t DIT1t DIT1t DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d2 NAB DIT1t-d2 NAB DIT1t-d2 NAB DIT1t-d2 NAB DIT1t-d2 PAP DIT1t-d2 PAP DIT1t-d2 PAP DIT1t-d2 PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d18 NAB DIT1t-d18 NAB DIT1t-d18 NAB DIT1t-d18 NAB DIT1t-d18 PAP DIT1t-d18 PAP DIT1t-d18 PAP DIT1t-d18 PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d19 NAB DIT1t-d19 NAB DIT1t-d19 NAB DIT1t-d19 NAB DIT1t-d19 PAP DIT1t-d19 PAP DIT1t-d19 PAP DIT1t-d19 PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d20 NAB DIT1t-d20 NAB DIT1t-d20 NAB DIT1t-d20 NAB DIT1t-d20 PAP DIT1t-d20 PAP DIT1t-d20 PAP DIT1t-d20 PAP DIT1t-d DIT1t-d DIT1t-d DIT1t-d DIT1t-d21 NAB DIT1t-d21 NAB DIT1t-d21 NAB DIT1t-d21 NAB DIT1t-d21 PAP DIT1t-d21 PAP DIT1t-d21 PAP DIT1t-d21 PAP
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