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Transcriptional activity of RNA polymerase II is modulated during the cell cycle. We previously identified a temperature-sensitive mutation in the largest (catalytic) subunit of RNA polymerase II (RPB1) that causes cell cycle arrest and genome instability. We now characterize a different cell line that has a temperature-sensitive defect in cell cycle progression, and find that it also has a mutation in RPB1. The temperature-sensitive mutant, tsAF8, of the Syrian hamster cell line, BHK21, arrests at the non-permissive temperature in the mid-G(1) phase. We show that RPB1 in tsAF8--which is found exclusively in the nucleus at the permissive temperature--is also found in the cytoplasm at the non-permissive temperature. Comparison of the DNA sequences of the RPB1 gene in the wild-type and mutant shows the mutant phenotype results from a (hemizygous) C-to-A variation at nucleotide 944 in one RPB1 allele; this gives rise to an ala-to-asp substitution at residue 315 in the protein. Aligning the amino acid sequences from various species reveals that ala(315) is highly conserved in eukaryotes.

Type

Journal article

Journal

Gene

Publication Date

22/08/2001

Volume

274

Pages

77 - 81

Keywords

Amino Acid Sequence, Animals, Base Sequence, Catalytic Domain, Cell Cycle, Cell Line, DNA, Complementary, G1 Phase, Genotype, Molecular Sequence Data, Mutation, Phylogeny, RNA Polymerase II, Sequence Alignment, Sequence Analysis, DNA, Sequence Homology, Amino Acid, Temperature