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PMID: 9528765 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Analysis of the interaction of the novel RNA polymerase II (pol II) subunit hsRPB4 with its partner hsRPB7 and with pol II.

Molecular and cellular biology ·Vol. 18 ·No. 4 ·1998-04-00 ·Pages 1935-45

Khazak V, Estojak J, Cho H, Majors J, Sonoda G, Testa JR, Golemis EA

Abstract

Under conditions of environmental stress, prokaryotes and lower eukaryotes such as the yeast Saccharomyces cerevisiae selectively utilize particular subunits of RNA polymerase II (pol II) to alter transcription to patterns favoring survival. In S. cerevisiae, a complex of two such subunits, RPB4 and RPB7, preferentially associates with pol II during stationary phase; of these two subunits, RPB4 is specifically required for survival under nonoptimal growth conditions. Previously, we have shown that RPB7 possesses an evolutionarily conserved human homolog, hsRPB7, which was capable of partially interacting with RPB4 and the yeast transcriptional apparatus. Using this as a probe in a two-hybrid screen, we have now established that hsRPB4 is also conserved in higher eukaryotes. In contrast to hsRPB7, hsRPB4 has diverged so that it no longer interacts with yeast RPB7, although it partially complements rpb4- phenotypes in yeast. However, hsRPB4 associates strongly and specifically with hsRPB7 when expressed in yeast or in mammalian cells and copurifies with intact pol II. hsRPB4 expression in humans parallels that of hsRPB7, supporting the idea that the two proteins may possess associated functions. Structure-function studies of hsRPB4-hsRPB7 are used to establish the interaction interface between the two proteins. This identification completes the set of human homologs for RNA pol II subunits defined in yeast and should provide the basis for subsequent structural and functional characterization of the pol II holoenzyme.

MeSH Terms
Amino Acid Sequence Animals COS Cells Cloning, Molecular HeLa Cells Humans Molecular Sequence Data Peptide Mapping Protein Binding Protein Conformation RNA Polymerase II/genetics,metabolism
Chemicals
RNA Polymerase II
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Khazak V
Division of Basic Sciences, Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111, USA.
Estojak J
Cho H
Majors J
Sonoda G
Testa J R
Golemis E A
References (67)
67 references, click to expand
  1. A 14.4 KDa acidic subunit of human RNA polymerase II with a putative leucine-zipper.
    DNA Seq. 1994;4(5):329-31 PMID: 7803819
  2. A kinase-cyclin pair in the RNA polymerase II holoenzyme.
    Nature. 1995 Mar 9;374(6518):193-6 PMID: 7877695
  3. RNA polymerase II subunit RPB9 is required for accurate start site selection.
    Genes Dev. 1995 Feb 15;9(4):481-90 PMID: 7883169
  4. High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier.
    Curr Genet. 1989 Dec;16(5-6):339-46 PMID: 2692852
  5. The amino acid sequence of the human RNA polymerase II 33-kDa subunit hRPB 33 is highly conserved among eukaryotes.
    J Biol Chem. 1990 May 25;265(15):8400-3 PMID: 2187864
  6. RNA polymerase B (II) and general transcription factors.
    Annu Rev Biochem. 1990;59:711-54 PMID: 2197989
  7. Mapping small DNA sequences by fluorescence in situ hybridization directly on banded metaphase chromosomes.
    Proc Natl Acad Sci U S A. 1990 Aug;87(16):6223-7 PMID: 2201023
  8. Basic local alignment search tool.
    J Mol Biol. 1990 Oct 5;215(3):403-10 PMID: 2231712
  9. RNA polymerase II: subunit structure and function.
    Trends Biochem Sci. 1990 Sep;15(9):347-51 PMID: 1700503
  10. Two dissociable subunits of yeast RNA polymerase II stimulate the initiation of transcription at a promoter in vitro.
    J Biol Chem. 1991 Jan 5;266(1):71-5 PMID: 1985924
  11. RNA polymerase II.
    Annu Rev Biochem. 1991;60:689-715 PMID: 1883205
  12. Yeast RNA polymerase II subunit RPB9 is essential for growth at temperature extremes.
    J Biol Chem. 1991 Oct 5;266(28):19053-5 PMID: 1918023
  13. The nonphosphorylated form of RNA polymerase II preferentially associates with the preinitiation complex.
    Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):10004-8 PMID: 1946417
  14. The nontransmembrane tyrosine phosphatase PTP-1B localizes to the endoplasmic reticulum via its 35 amino acid C-terminal sequence.
    Cell. 1992 Feb 7;68(3):545-60 PMID: 1739967
  15. Complete sequence of the human RNA polymerase II largest subunit.
    Nucleic Acids Res. 1992 Feb 25;20(4):910 PMID: 1542581
  16. RPC10 encodes a new mini subunit shared by yeast nuclear RNA polymerases.
    Gene Expr. 1992;2(1):31-7 PMID: 1617300
  17. Fused protein domains inhibit DNA binding by LexA.
    Mol Cell Biol. 1992 Jul;12(7):3006-14 PMID: 1620111
  18. Chance and statistical significance in protein and DNA sequence analysis.
    Science. 1992 Jul 3;257(5066):39-49 PMID: 1621093
  19. Primary structure of the second largest subunit of human RNA polymerase II (or B).
    J Mol Biol. 1992 Aug 20;226(4):1295-9 PMID: 1518060
  20. Cytostellin: a novel, highly conserved protein that undergoes continuous redistribution during the cell cycle.
    J Cell Sci. 1992 Oct;103 ( Pt 2):381-8 PMID: 1478941
  21. Survival of hunger and stress: the role of rpoS in early stationary phase gene regulation in E. coli.
    Cell. 1993 Jan 29;72(2):165-8 PMID: 8425216
  22. Heterogeneity of the principal sigma factor in Escherichia coli: the rpoS gene product, sigma 38, is a second principal sigma factor of RNA polymerase in stationary-phase Escherichia coli.
    Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3511-5 PMID: 8475100
  23. Protein localization and asymmetry in the bacterial cell.
    Cell. 1993 Jun 4;73(5):841-55 PMID: 8500178
  24. General initiation factors for RNA polymerase II.
    Annu Rev Biochem. 1993;62:161-90 PMID: 8352588
  25. A growth rate-limiting process in the last growth phase of the yeast life cycle involves RPB4, a subunit of RNA polymerase II.
    J Bacteriol. 1993 Oct;175(19):6358-63 PMID: 8407810
  26. A portion of RNA polymerase II molecules has a component essential for stress responses and stress survival.
    Mol Cell Biol. 1993 Nov;13(11):6984-91 PMID: 8413288
  27. Cdi1, a human G1 and S phase protein phosphatase that associates with Cdk2.
    Cell. 1993 Nov 19;75(4):791-803 PMID: 8242750
  28. Tyrosine phosphorylation of mammalian RNA polymerase II carboxyl-terminal domain.
    Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11167-71 PMID: 7504297
  29. Association of Krev-1/rap1a with Krit1, a novel ankyrin repeat-containing protein encoded by a gene mapping to 7q21-22.
    Oncogene. 1997 Aug 28;15(9):1043-9 PMID: 9285558
  30. Two conformations of RNA polymerase II revealed by electron crystallography.
    J Mol Biol. 1997 Oct 3;272(4):536-40 PMID: 9325110
  31. Structure of wild-type yeast RNA polymerase II and location of Rpb4 and Rpb7.
    EMBO J. 1998 Apr 15;17(8):2353-8 PMID: 9545247
  32. The RNA polymerase II holoenzyme and its implications for gene regulation.
    Trends Biochem Sci. 1995 Mar;20(3):113-6 PMID: 7709429
  33. Transcription-dependent redistribution of the large subunit of RNA polymerase II to discrete nuclear domains.
    J Cell Biol. 1995 Apr;129(2):287-98 PMID: 7536746
  34. Chromosomal localization of a gene, GF1, encoding a novel zinc finger protein reveals a new syntenic region between man and rodents.
    Cytogenet Cell Genet. 1995;70(3-4):263-7 PMID: 7789186
  35. Four subunits that are shared by the three classes of RNA polymerase are functionally interchangeable between Homo sapiens and Saccharomyces cerevisiae.
    Mol Cell Biol. 1995 Sep;15(9):4702-10 PMID: 7651387
  36. Correlation of two-hybrid affinity data with in vitro measurements.
    Mol Cell Biol. 1995 Oct;15(10):5820-9 PMID: 7565735
  37. Short transcripts of the ternary complex provide insight into RNA polymerase II elongational pausing.
    J Mol Biol. 1995 Oct 6;252(5):522-35 PMID: 7563071
  38. Initial assessment of human gene diversity and expression patterns based upon 83 million nucleotides of cDNA sequence.
    Nature. 1995 Sep 28;377(6547 Suppl):3-174 PMID: 7566098
  39. Six human RNA polymerase subunits functionally substitute for their yeast counterparts.
    Mol Cell Biol. 1995 Dec;15(12):6895-900 PMID: 8524256
  40. Human RNA polymerase II subunit hsRPB7 functions in yeast and influences stress survival and cell morphology.
    Mol Biol Cell. 1995 Jul;6(7):759-75 PMID: 7579693
  41. A human RNA polymerase II complex associated with SRB and DNA-repair proteins.
    Nature. 1996 May 2;381(6577):86-9 PMID: 8609996
  42. Human enhancer of filamentation 1, a novel p130cas-like docking protein, associates with focal adhesion kinase and induces pseudohyphal growth in Saccharomyces cerevisiae.
    Mol Cell Biol. 1996 Jul;16(7):3327-37 PMID: 8668148
  43. A hyperphosphorylated form of the large subunit of RNA polymerase II is associated with splicing complexes and the nuclear matrix.
    Proc Natl Acad Sci U S A. 1996 Aug 6;93(16):8253-7 PMID: 8710856
  44. The gene (POLR2L) encoding the hRPB7.6 subunit of human RNA polymerase.
    Genomics. 1996 Feb 15;32(1):86-90 PMID: 8786124
  45. Genomic structure of the RNA polymerase II small subunit (hRPB14.4) locus (POLRF) and mapping to 22q13.1 by sequence identity.
    Genomics. 1996 Jun 15;34(3):440-2 PMID: 8786150
  46. Cloning of a novel human RNA polymerase II subunit downregulated by doxorubicin: new potential mechanisms of drug related toxicity.
    FEBS Lett. 1996 Apr 8;384(1):48-52 PMID: 8797801
  47. Comparative analysis of 1196 orthologous mouse and human full-length mRNA and protein sequences.
    Genome Res. 1996 Sep;6(9):846-57 PMID: 8889551
  48. Purification of human RNA polymerase II and general transcription factors.
    Methods Enzymol. 1996;274:72-100 PMID: 8902797
  49. Transcriptional activation by recruitment.
    Nature. 1997 Apr 10;386(6625):569-77 PMID: 9121580
  50. Transcription elongation through DNA arrest sites. A multistep process involving both RNA polymerase II subunit RPB9 and TFIIS.
    J Biol Chem. 1997 Jun 6;272(23):14747-54 PMID: 9169440
  51. An electron-transport system associated with the outer membrane of liver mitochondria. A biochemical and morphological study.
    J Cell Biol. 1967 Feb;32(2):415-38 PMID: 10976232
  52. Preparation and properties of plasma membrane and endoplasmic reticulum fragments from isolated rat fat cells.
    Biochim Biophys Acta. 1971 Apr 13;233(2):334-47 PMID: 4326970
  53. Animal DNA-dependent RNA polymerases. Molecular structures and immunological properties of calf-thymus enzyme AI and of calf-thymus and rat-liver enzymes B.
    Eur J Biochem. 1974 May 15;44(2):421-36 PMID: 4209621
  54. Purification using polyethylenimine precipitation and low molecular weight subunit analyses of calf thymus and wheat germ DNA-dependent RNA polymerase II.
    Biochemistry. 1977 May 31;16(11):2334-43 PMID: 558798
  55. The lexA gene product represses its own promoter.
    Proc Natl Acad Sci U S A. 1980 Apr;77(4):1932-6 PMID: 6990417
  56. A simple, rapid, and sensitive DNA assay procedure.
    Anal Biochem. 1980 Mar 1;102(2):344-52 PMID: 6158890
  57. Eukaryotic RNA polymerases.
    CRC Crit Rev Biochem. 1985;18(1):31-90 PMID: 3893883
  58. Characterization of RNA polymerase type II from human term placenta.
    J Cell Physiol. 1986 Jun;127(3):432-8 PMID: 3754875
  59. Factors involved in specific transcription by mammalian RNA polymerase II. Purification and functional analysis of initiation factors IIB and IIE.
    J Biol Chem. 1987 Mar 5;262(7):3310-21 PMID: 3029109
  60. Isolation and molecular characterization of a cDNA encoding the 23-kDa subunit of human RNA polymerase II.
    J Biol Chem. 1989 Aug 5;264(22):13114-21 PMID: 2753903
  61. RNA polymerase II subunit RPB4 is essential for high- and low-temperature yeast cell growth.
    Mol Cell Biol. 1989 Jul;9(7):2854-9 PMID: 2674672
  62. Structure of the gene encoding the 14.5 kDa subunit of human RNA polymerase II.
    Nucleic Acids Res. 1993 Nov 25;21(23):5345-50 PMID: 8265347
  63. The basics of basal transcription by RNA polymerase II.
    Cell. 1994 Apr 8;77(1):1-3 PMID: 8156586
  64. Transcriptional activation: a complex puzzle with few easy pieces.
    Cell. 1994 Apr 8;77(1):5-8 PMID: 8156597
  65. Functional substitution of an essential yeast RNA polymerase subunit by a highly conserved mammalian counterpart.
    Mol Cell Biol. 1994 Jun;14(6):4155-9 PMID: 8196653
  66. Applications of statistical criteria in protein sequence analysis: case study of yeast RNA polymerase II subunits.
    Comput Chem. 1994 Sep;18(3):251-3 PMID: 7952895
  67. Relationship of CDK-activating kinase and RNA polymerase II CTD kinase TFIIH/TFIIK.
    Cell. 1994 Dec 16;79(6):1103-9 PMID: 8001136
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.18.4.1935
Published
1998-04-00
Pages
1935-45
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC121423
Subset
IM
Grants
NCI NIH HHS · CA-06927 · United States
NCI NIH HHS · CA-70841 · United States
Databases
GENBANK
U85510, U89387
External Links
PubMed source DOI: 10.1128/MCB.18.4.1935 PMC Full Text View journal details
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