Mouse primase p49 subunit molecular cloning indicates conserved and divergent regions.
Primase is a specialized RNA polymerase that synthesizes RNA primers for initiation of DNA synthesis. A full cDNA clone of the p49 subunit of mouse primase, a heterodimeric enzyme, has been isolated using a primase p49-specific polyclonal antibody to screen a lambda gt11 mouse cDNA expression library. The cDNA indicated the subunit is a 417-amino acid polypeptide with a calculated molecular mass of 49,295 daltons. The p49 mRNA is approximately 1500 nucleotides long with a 5'-untranslated region of 74 nucleotides and a 3'-untranslated region of 200 nucleotides. Comparison with a similar sized primase subunit from yeast showed highly conserved amino acid sequences in the N-terminal halves of the polypeptides and included a potential metal-binding domain suggesting the functional importance of this region for DNA binding. In contrast, the 3' portion of the cDNA has rapidly diverged in nucleotide sequence, as primase mRNA can be detected in mouse and rat cells with a 3' probe (including coding and noncoding) but not in RNA from hamster or human cells. A full-length cDNA probe detected mRNA from hamster and human cell lines, indicating a conserved 5' portion and divergent 3' region of the expressed gene. The rapid divergence may be related to the species-specific protein interactions found for the DNA polymerase alpha-primase complex. The mRNA is detected in proliferating but not in quiescent cells consistent with its function in DNA replication.
Prussak
C E
CE
Department of Medicine, University of California, San Diego, La Jolla 92093.
Almazan
M T
MT
Tseng
B Y
BY
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D006224:Cricetinae; D004247:DNA / Q000302:isolation & purification; D019915:DNA Primase; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000302:isolation & purification; D014176:Protein Biosynthesis; D012316:RNA Nucleotidyltransferases / Q000235:genetics* / Q000302:isolation & purification; D011817:Rabbits; D051381:Rats; D012689:Sequence Homology, Nucleic Acid
D010446:Peptide Fragments; D004247:DNA; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases