Isolation and molecular characterization of a cDNA encoding the 23-kDa subunit of human RNA polymerase II.
We have shown that antibodies against native calf thymus RNA polymerase II and antibodies against its 23-kDa subunit cross-reacted with the 23-kDa subunit of human RNA polymerase II. Immunoglobin G (IgG) against the 23-kDa subunit of calf thymus RNA polymerase II inhibited transcription in vitro from the adenovirus major late promoter. By immunoscreening of a human placenta lambda gt11 cDNA library with IgG against native CT RNA polymerase II and with IgG against its 23-kDa subunit, we isolated and characterized a full length 1.2-kilobase cDNA. We also generated oligonucleotide probes from a sequence of amino acid residues obtained by a modified peptide microsequencing procedure. The cDNAs isolated both from oligoscreening and immunoscreening were identical. The amino acid sequence deduced from the nucleotide sequence analysis indicates a polypeptide of 197 amino acid (23 kDa). The in vitro translation product of human cDNA HP-23 was precipitated by IgG against the 23-kDa subunit of CT RNA polymerase II. The amino acid sequence deduced from HP-23 showed no obvious homology with Escherichia coli RNA polymerase subunits or with any of its sigma factors.
Pati
U K
UK
Department of Human Genetics, Yale University School of Medicine, New Haven, Connecticut 06510.
Weissman
S M
SM
D000256:Adenoviridae / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002417:Cattle; D003001:Cloning, Molecular; D003429:Cross Reactions; D004247:DNA / Q000302:isolation & purification*; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D007074:Immunoglobulin G / Q000502:physiology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D012319:RNA Polymerase II / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D013950:Thymus Gland / Q000201:enzymology; D014158:Transcription, Genetic
D007074:Immunoglobulin G; D004247:DNA; D012319:RNA Polymerase II