Isolation and characterization of cDNA encoding mouse RNA polymerase II subunit RPB14

• Published in: Gene 1997-03, Vol.187 (2), p.165-170
• Main Authors: Nishi, Yuichi, Yamamoto, Kazuo, Yao, Yeqi, Yamamoto, Mika, Nogi, Yasuhisa, Matsuo, Hiroshi, Muramatsu, Masami
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Binding Sites; DNA, Complementary; Humans; Mice; Molecular Sequence Data; Protein-protein interaction; RNA Polymerase II - genetics; Sequence Homology, Amino Acid; Two-hybrid system; α-Motif
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/S0378-1119(96)00739-1

By means of the yeast two-hybrid system using the 40-kDa subunit of mouse RNA polymerase I, mRPA40, as the bait, we isolated a mouse cDNA which encoded a protein with significant homology in amino acid sequence to the 12.5-kDa subunit of Saccharomyces cerevisiae RNA polymerase II, B12.5 (RPB11). Specific antibody raised against the recombinant protein that was derived from the cDNA reacted with a 14-kDa polypeptide in highly purified mammalian RNA polymerase II and did not react with any subunit of RNA polymerase I or III. Moreover, the antibody co-immunoprecipitated the largest subunit of mouse RNA polymerase II. These results provide biochemical evidence that the cDNA isolated, named mRPB14, encodes a specific subunit of RNA polymerase II, and indicate that the subunit organization of the enzyme is conserved between yeast and mouse. A possible role of the α-motif [Dequard-Chablat, M., Riva, M., Carles, C. and Sentenac, A., J. Biol. Chem. 266 (1991) 15300–15307] in the protein-protein interaction between mRPA40 and mRPB14 is also discussed.