Abstract
Both the gene and the cDNA encoding the Rpb4 subunit of RNA polymerase II were cloned from the fission yeast Schizosaccharomyces pombe. The cDNA sequence indicates that Rpb4 consists of 135 amino acid residues with a molecular weight of 15,362. As in the case of the corresponding subunits from higher eukaryotes such as humans and the plant Arabidopsis thaliana, Rpb4 is smaller than RPB4 from the budding yeast Saccharomyces cerevisiae and lacks several segments, which are present in the S. cerevisiae RPB4 subunit, including the highly charged sequence in the central portion. The RPB4 subunit of S. cerevisiae is not essential for normal cell growth but is required for cell viability under stress conditions. In contrast, S. pombe Rpb4 was found to be essential even under normal growth conditions. The fraction of RNA polymerase II containing RPB4 in exponentially growing cells of S. cerevisiae is about 20%, but S. pombe RNA polymerase II contains the stoichiometric amount of Rpb4 even at the exponential growth phase. In contrast to the RPB4 homologues from higher eukaryotes, however, S. pombe Rpb4 formed stable hybrid heterodimers with S. cerevisiae RPB7, suggesting that S. pombe Rpb4 is similar, in its structure and essential role in cell viability, to the corresponding subunits from higher eukaryotes. However, S. pombe Rpb4 is closer in certain molecular functions to S. cerevisiae RPB4 than the eukaryotic RPB4 homologues.
MeSH Terms
Amino Acid Sequence
Cell Division/genetics
Cloning, Molecular
DNA, Complementary/genetics
Dimerization
Eukaryotic Cells
Fungal Proteins/metabolism
Genes, Essential
Genes, Fungal
Molecular Sequence Data
Mutagenesis
Protein Binding
Protein Conformation
RNA Polymerase II/chemistry,genetics,metabolism
Schizosaccharomyces/enzymology,genetics
Sequence Analysis, DNA
Sequence Homology, Amino Acid
Chemicals
DNA, Complementary
Fungal Proteins
RNA Polymerase II
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Sakurai H
Department of Molecular Genetics, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
Mitsuzawa H
Kimura M
Ishihama A
References (27)
27 references, click to expand
-
Involvement of multiple subunit-subunit contacts in the assembly of RNA polymerase II.
Nucleic Acids Res. 2000 Feb 15;28(4):952-9
PMID: 10648788
-
Two forms of RNA polymerase B in yeast. Proteolytic conversion in vitro of enzyme BI into BII.
Eur J Biochem. 1976 Jun 1;65(2):543-52
PMID: 780108
-
A mutation of the B220 subunit gene affects the structural and functional properties of yeast RNA polymerase B in vitro.
J Biol Chem. 1980 Jul 10;255(13):6450-5
PMID: 6993472
-
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
-
RNA polymerase II subunit composition, stoichiometry, and phosphorylation.
Mol Cell Biol. 1990 May;10(5):1915-20
PMID: 2183013
-
High-frequency transformation method and library transducing vectors for cloning mammalian cDNAs by trans-complementation of Schizosaccharomyces pombe.
Nucleic Acids Res. 1990 Nov 25;18(22):6485-9
PMID: 2251111
-
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
-
Molecular genetic analysis of fission yeast Schizosaccharomyces pombe.
Methods Enzymol. 1991;194:795-823
PMID: 2005825
-
RNA polymerase II.
Annu Rev Biochem. 1991;60:689-715
PMID: 1883205
-
Subunits of the Schizosaccharomyces pombe RNA polymerase II: enzyme purification and structure of the subunit 3 gene.
Nucleic Acids Res. 1993 Aug 11;21(16):3749-54
PMID: 8367291
-
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
-
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
-
Fission yeast gene structure and recognition.
Nucleic Acids Res. 1994 May 11;22(9):1750-9
PMID: 8202381
-
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
-
Subunit composition of RNA polymerase II from the fission yeast Schizosaccharomyces pombe.
Gene. 1996 Nov 21;180(1-2):63-7
PMID: 8973348
-
Gene organization and protein sequence of the small subunits of Schizosaccharomyces pombe RNA polymerase II.
Gene. 1997 Sep 1;196(1-2):165-74
PMID: 9322754
-
Two conformations of RNA polymerase II revealed by electron crystallography.
J Mol Biol. 1997 Oct 3;272(4):536-40
PMID: 9325110
-
RNA polymerase II subunits 2, 3, and 11 form a core subassembly with DNA binding activity.
J Biol Chem. 1997 Oct 10;272(41):25851-5
PMID: 9325316
-
Construction and characterization of a full length-enriched and a 5'-end-enriched cDNA library.
Gene. 1997 Oct 24;200(1-2):149-56
PMID: 9373149
-
Two small subunits in Arabidopsis RNA polymerase II are related to yeast RPB4 and RPB7 and interact with one another.
J Biol Chem. 1998 Mar 6;273(10):5631-7
PMID: 9488692
-
Analysis of the interaction of the novel RNA polymerase II (pol II) subunit hsRPB4 with its partner hsRPB7 and with pol II.
Mol Cell Biol. 1998 Apr;18(4):1935-45
PMID: 9528765
-
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
-
Two large subunits of the fission yeast RNA polymerase II provide platforms for the assembly of small subunits.
J Mol Biol. 1998 Jun 19;279(4):703-12
PMID: 9642054
-
Rpb4, a subunit of RNA polymerase II, enables the enzyme to transcribe at temperature extremes in vitro.
J Bacteriol. 1998 Dec;180(23):6187-92
PMID: 9829926
-
Identification of the gene and the protein of RNA polymerase II subunit 9 (Rpb9) from the fission yeast Schizosacharomyces pombe.
Gene. 1998 Oct 9;221(1):11-6
PMID: 9852944
-
Subunits of yeast RNA polymerases: structure and function.
Curr Opin Microbiol. 1998 Apr;1(2):190-6
PMID: 10066472
-
Arabidopsis thaliana RNA polymerase II subunits related to yeast and human RPB5.
Gene. 1999 Apr 29;231(1-2):41-7
PMID: 10231567