Molecular Structure of Yeast RNA Polymerase III: Demonstration of the Tripartite Transcriptive System in Lower Eukaryotes

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1976 Apr 1

PMID 772675

Citations 24

Authors

P Valenzuela

G L Hager

F Weinberg

W J Rutter

Affiliations

Soon will be listed here.

Abstract

Homogeneous RNA polymerase III (RNA nucleotidyltransferase III) has been obtained from yeast. The subunit composition of the enzyme was examined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The enzyme is composed of 12 putative subunits with molecular weights 160,000, 128,000, 82,000, 41,000, 40,500, 37,000, 34,000, 28,000, 24,000, 20,000, 14,500, and 11,000. The high-molecular-weight subunits and several of the smaller subunits of yeast RNA polymerase III are clearly different from those of enzymes I and II, indicating a distinct molecular structure. However, the molecular weights of some of the small subunits (41,000, 28,000, 24,000, and 14,500) appear to be identical to those of polymerases I and II. Thus, it is possible that the three classes of enzymes in yeast have some common subunits. As in other eukaryotes, yeast polymerase II is inhibited by relatively low concentrations of alpha-amanitin; however, contrary to what has been found in higher eukaryotes, yeast polymerase III is resistant (up to 2 mg/ml) to alpha-amanitin, while yeast polymerase I is sensitive to high concentrations of the drug (50% inhibition at 0.3 mg/ml). These results establish the existence of RNA polymerase III in yeast and provide a structural basis for the discrimination of the three functional polymerases in eukaryotes.

Citing Articles

Dynamic structures of intrinsically disordered proteins related to the general transcription factor TFIIH, nucleosomes, and histone chaperones.

Okuda M, Tsunaka Y, Nishimura Y Biophys Rev. 2023; 14(6):1449-1472.

PMID: 36659983 PMC: 9842849. DOI: 10.1007/s12551-022-01014-9.

Structure-function analysis of hRPC62 provides insights into RNA polymerase III transcription initiation.

Lefevre S, Dumay-Odelot H, El-Ayoubi L, Budd A, Legrand P, Pinaud N Nat Struct Mol Biol. 2011; 18(3):352-8.

PMID: 21358628 DOI: 10.1038/nsmb.1996.

Cell growth- and differentiation-dependent regulation of RNA polymerase III transcription.

Dumay-Odelot H, Durrieu-Gaillard S, da Silva D, Roeder R, Teichmann M Cell Cycle. 2010; 9(18):3687-99.

PMID: 20890107 PMC: 3047797. DOI: 10.4161/cc.9.18.13203.

Recruitment of RNA polymerase III in vivo.

Kenneth N, Marshall L, White R Nucleic Acids Res. 2008; 36(11):3757-64.

PMID: 18487626 PMC: 2441808. DOI: 10.1093/nar/gkn272.

Novel small-molecule inhibitors of RNA polymerase III.

Wu L, Pan J, Thoroddsen V, Wysong D, Blackman R, Bulawa C Eukaryot Cell. 2003; 2(2):256-64.

PMID: 12684375 PMC: 154847. DOI: 10.1128/EC.2.2.256-264.2003.

References

Roeder R, Rutter W . Multiple forms of DNA-dependent RNA polymerase in eukaryotic organisms. Nature. 1969; 224(5216):234-7. DOI: 10.1038/224234a0. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Chambon P . Eukaryotic nuclear RNA polymerases. Annu Rev Biochem. 1975; 44:613-38. DOI: 10.1146/annurev.bi.44.070175.003145. View

Weinmann R, Raskas H, Roeder R . Role of DNA-dependent RNA polymerases II and III in transcription of the adenovirus genome late in productive infection. Proc Natl Acad Sci U S A. 1974; 71(9):3426-39. PMC: 433786. DOI: 10.1073/pnas.71.9.3426. View

Sklar V, Schwartz L, Roeder R . Distinct molecular structures of nuclear class I, II, and III DNA-dependent RNA polymerases. Proc Natl Acad Sci U S A. 1975; 72(1):348-52. PMC: 432302. DOI: 10.1073/pnas.72.1.348. View

Schultz L, Hall B . Transcription in yeast: alpha-amanitin sensitivity and other properties which distinguish between RNA polymerases I and III. Proc Natl Acad Sci U S A. 1976; 73(4):1029-33. PMC: 430193. DOI: 10.1073/pnas.73.4.1029. View

Schwartz L, Sklar V, Jaehning J, Weinmann R, Roeder R . Isolation and partial characterization of the multiple forms of deoxyribonucleic acid-dependent ribonucleic acid polymerase in the mouse myeloma, MOPC 315. J Biol Chem. 1974; 249(18):5889-97. View

Weber K, Osborn M . The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969; 244(16):4406-12. View

Ullmann A, Goldberg M, Perrin D, MONOD J . On the determination of molecular weight of proteins and protein subunits in the presence of 6 M guanidine hydrochloride. Biochemistry. 1968; 7(1):261-5. DOI: 10.1021/bi00841a031. View

10.

Brogt T, Planta R . Characteristics of DNA-dependent RNA polymerase activity from isolated yeast nuclei. FEBS Lett. 1972; 20(1):47-52. DOI: 10.1016/0014-5793(72)80014-0. View

11.

Buhler J, Sentenac A, FROMAGEOT P . Isolation, structure, and general properties of yeast ribonucleic acid polymerase A (or I). J Biol Chem. 1974; 249(18):5963-70. View

12.

Armstrong J, MYERS D, VERPOORTE J, EDSALL J . Purification and properties of human erythrocyte carbonic anhydrases. J Biol Chem. 1966; 241(21):5137-49. View

13.

Ponta H, Ponta U, Wintersberger E . DNA-dependent RNA polymerases from yeast. Partial characterization of three nuclear enzyme activities. FEBS Lett. 1971; 18(2):204-208. DOI: 10.1016/0014-5793(71)80445-3. View

14.

Burgess R . Separation and characterization of the subunits of ribonucleic acid polymerase. J Biol Chem. 1969; 244(22):6168-76. View

15.

Adman R, Schultz L, Hall B . Transcription in yeast: separation and properties of multiple FNA polymerases. Proc Natl Acad Sci U S A. 1972; 69(7):1702-6. PMC: 426782. DOI: 10.1073/pnas.69.7.1702. View

16.

Weinmann R, Roeder R . Role of DNA-dependent RNA polymerase 3 in the transcription of the tRNA and 5S RNA genes. Proc Natl Acad Sci U S A. 1974; 71(5):1790-4. PMC: 388326. DOI: 10.1073/pnas.71.5.1790. View

17.

Lindell T, Weinberg F, MORRIS P, Roeder R, Rutter W . Specific inhibition of nuclear RNA polymerase II by alpha-amanitin. Science. 1970; 170(3956):447-9. DOI: 10.1126/science.170.3956.447. View

18.

Gornicki S, Vuturo S, West T, Weaver R . Purification and properties of deoxyribonucleic acid-dependent ribonucleic acid polymerases from the slime mold Physarum polycephalum. J Biol Chem. 1974; 249(6):1792-8. View

19.

Pong S, LOOMIS Jr W . Multiple nuclear ribonucleic acid polymerases during development of Dictyostelium discoideum. J Biol Chem. 1973; 248(11):3933-9. View

20.

Huet J, Buhler J, Sentenac A, FROMAGEOT P . Dissociation of two polypeptide chains from yeast RNA polymerase A. Proc Natl Acad Sci U S A. 1975; 72(8):3034-8. PMC: 432913. DOI: 10.1073/pnas.72.8.3034. View