Domains of Escherichia coli primase: functional activity of a 47-kDa N-terminal proteolytic fragment.
Endoproteinase Asp-N cleaves the 581-amino acid Escherichia coli primase (65,564 Da) into several major fragments. One of these, a 47-kDa fragment containing the complete N terminus and the first 422 amino acids of primase, is capable of primer RNA (pRNA) synthesis in the G4oric/single-stranded DNA binding protein/primase pRNA synthesis system. A cloned 398-amino acid N-terminal fragment of primase can also synthesize pRNA. The sizes of the pRNA synthesized by these N-terminal fragments, however, are smaller than those synthesized by intact primase, suggesting that the C-terminal region of primase plays a role in processivity or regulation of pRNA synthesis. Primase mutants with the last 10 and 40 C-terminal amino acids deleted synthesize pRNA as wild-type primase, indicating that any regulatory sequences must be internal to the C terminus of primase.
Sun
W
W
Biochemistry Department, New York University Medical Center, NY 10016.
Tormo
J
J
Steitz
T A
TA
Godson
G N
GN
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D019915:DNA Primase; D004926:Escherichia coli / Q000201:enzymology*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D010446:Peptide Fragments / Q000378:metabolism; D012316:RNA Nucleotidyltransferases / Q000737:chemistry*; D012329:RNA, Bacterial / Q000096:biosynthesis; D013329:Structure-Activity Relationship
D009838:Oligodeoxyribonucleotides; D010446:Peptide Fragments; D012329:RNA, Bacterial; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases