A Selection for Mutants of the RNA Polymerase III Transcription Apparatus: PCF1 Stimulates Transcription of TRNA and 5S RNA Genes

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1989 Dec 20

PMID 2686985

Citations 15

Authors

I WILLIS

P Schmidt

D Soll

Affiliations

Soon will be listed here.

Abstract

A genetic approach has been developed to study transcription by RNA polymerase III. A pair of Schizosaccharomyces pombe nonsense suppressor tRNA genes were arranged in tandem such that expression of the downstream (supS1) tRNA suppressor was dependent upon transcription initiated by the internal promoter of the upstream (sup9-e) gene. Dominant mutant strains of Saccharomyces cerevisiae were isolated that suppress in trans the effect of an A block promoter mutation (A19) in the sup9-e gene and restore supS1 suppressor activity. Fifteen mutant strains, eight of which were independently isolated, all have elevated steady-state levels of sup9-e A19 RNA consistent with an increase in gene transcription. Extracts of a strain carrying the dominant mutant gene, PCF1, show a general 6-fold stimulation in transcription of mutant (A19) and wild-type tRNA genes and increase 5S gene transcription 4-fold compared with extracts from a wild-type strain. A transcription factor exclusion assay was used to show that the PCF1 mutation affects two distinct stages in transcription: one prior to and one after stable complex formation; and that these effects are mediated by a component of the stable complex. Further evidence of an effect during complex assembly was obtained in a time-course experiment that showed a shortened lag phase in the PCF1 extract. The results indicate that PCF1 is either a component of the stable complex or a positive regulator of its activity.

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References

Klekamp M, Weil P . Specific transcription of homologous class III genes in yeast-soluble cell-free extracts. J Biol Chem. 1982; 257(14):8432-41. View

Kassavetis G, Riggs D, Negri R, Nguyen L, Geiduschek E . Transcription factor IIIB generates extended DNA interactions in RNA polymerase III transcription complexes on tRNA genes. Mol Cell Biol. 1989; 9(6):2551-66. PMC: 362328. DOI: 10.1128/mcb.9.6.2551-2566.1989. View

Allison D, Goh S, Hall B . The promoter sequence of a yeast tRNAtyr gene. Cell. 1983; 34(2):655-64. DOI: 10.1016/0092-8674(83)90398-7. View

Ruet A, Camier S, Smagowicz W, Sentenac A, FROMAGEOT P . Isolation of a class C transcription factor which forms a stable complex with tRNA genes. EMBO J. 1984; 3(2):343-50. PMC: 557347. DOI: 10.1002/j.1460-2075.1984.tb01809.x. View

Hottinger H, Stadelmann B, Pearson D, Frendewey D, Kohli J, Soll D . The Schizosaccharomyces pombe sup3-i suppressor recognizes ochre, but not amber codons in vitro and in vivo. EMBO J. 1984; 3(2):423-8. PMC: 557361. DOI: 10.1002/j.1460-2075.1984.tb01823.x. View

WILLIS I, Hottinger H, Pearson D, Chisholm V, Leupold U, Soll D . Mutations affecting excision of the intron from a eukaryotic dimeric tRNA precursor. EMBO J. 1984; 3(7):1573-80. PMC: 557561. DOI: 10.1002/j.1460-2075.1984.tb02013.x. View

Bieker J, Martin P, Roeder R . Formation of a rate-limiting intermediate in 5S RNA gene transcription. Cell. 1985; 40(1):119-27. DOI: 10.1016/0092-8674(85)90315-0. View

Baker R, Hall B . Structural features of yeast tRNA genes which affect transcription factor binding. EMBO J. 1984; 3(12):2793-800. PMC: 557768. DOI: 10.1002/j.1460-2075.1984.tb02211.x. View

Pearson D, WILLIS I, Hottinger H, Bell J, Kumar A, Leupold U . Mutations preventing expression of sup3 tRNASer nonsense suppressors of Schizosaccharomyces pombe. Mol Cell Biol. 1985; 5(4):808-15. PMC: 366785. DOI: 10.1128/mcb.5.4.808-815.1985. View

10.

Hoeffler W, Roeder R . Enhancement of RNA polymerase III transcription by the E1A gene product of adenovirus. Cell. 1985; 41(3):955-63. DOI: 10.1016/s0092-8674(85)80076-3. View

11.

Camier S, Gabrielsen O, Baker R, Sentenac A . A split binding site for transcription factor tau on the tRNA3Glu gene. EMBO J. 1985; 4(2):491-500. PMC: 554212. DOI: 10.1002/j.1460-2075.1985.tb03655.x. View

12.

Gaynor R, Feldman L, Berk A . Transcription of class III genes activated by viral immediate early proteins. Science. 1985; 230(4724):447-50. DOI: 10.1126/science.2996135. View

13.

Stillman D, Better M, Geiduschek E . Electron-microscopic examination of the binding of a large RNA polymerase III transcription factor to a tRNA gene. J Mol Biol. 1985; 185(2):451-5. DOI: 10.1016/0022-2836(85)90417-6. View

14.

Krupp G, Thurianx P, WILLIS I, Gamulin V, Soll D . First identification of an amber nonsense mutation in Schizosaccharomyces pombe: major differences in the efficiency of homologous versus heterologous yeast suppressor tRNA genes. Mol Gen Genet. 1985; 201(1):82-7. DOI: 10.1007/BF00397990. View

15.

Sharp S, Schaack J, Cooley L, Burke D, Soll D . Structure and transcription of eukaryotic tRNA genes. CRC Crit Rev Biochem. 1985; 19(2):107-44. DOI: 10.3109/10409238509082541. View

16.

Schaack J, Lapointe J, Mao J, Nichols M, Soll D . Dimeric tRNA gene arrangement in Schizosaccharomyces pombe allows increased expression of the downstream gene. Nucleic Acids Res. 1985; 13(24):8739-47. PMC: 318948. DOI: 10.1093/nar/13.24.8739. View

17.

Klekamp M, Weil P . Partial purification and characterization of the Saccharomyces cerevisiae transcription factor TFIIIB. J Biol Chem. 1986; 261(6):2819-27. View

18.

Baker R, Gabrielsen O, Hall B . Effects of tRNATyr point mutations on the binding of yeast RNA polymerase III transcription factor C. J Biol Chem. 1986; 261(12):5275-82. View

19.

WILLIS I, Frendewey D, Nichols M, Schaack J, Soll D . A single base change in the intron of a serine tRNA affects the rate of RNase P cleavage in vitro and suppressor activity in vivo in Saccharomyces cerevisiae. J Biol Chem. 1986; 261(13):5878-85. View

20.

WILLIS I, Nichols M, Chisholm V, Soll D, Heyer W, Szankasi P . Functional complementation between mutations in a yeast suppressor tRNA gene reveals potential for evolution of tRNA sequences. Proc Natl Acad Sci U S A. 1986; 83(20):7860-4. PMC: 386822. DOI: 10.1073/pnas.83.20.7860. View