Mouse DNA Polymerase α -primase Terminates and Reinitiates DNA Synthesis 2-14 Nucleotides Upstream of C2A1-2(C2-3/T<l atex>$_{2}$ ) Sequences on a Minute Virus of Mice DNA Template

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 82; no. 12; pp. 4023 - 4027
• Main Authors: Faust, Emanuel A., Nagy, Randy, Davey, Scott K.
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01.06.1985; National Acad Sciences
• Subjects: Animals; Base Sequence; Biochemistry; Biological and medical sciences; Chromosome Mapping; DNA; DNA Polymerase II - analysis; DNA Polymerase II - genetics; DNA Primase; DNA Replication; DNA, Viral - biosynthesis; DNA, Viral - genetics; Electrophoresis; Enzymes; Fundamental and applied biological sciences. Psychology; Gels; Genes, Regulator; KB cells; Macromolecular Substances; Mice; Minute virus of mice; Molecular and cellular biology; Molecular genetics; Molecular Weight; Nucleotides; Oligoribonucleotides; Replication; RNA; RNA Nucleotidyltransferases - analysis; RNA Nucleotidyltransferases - genetics; RNA, Viral - biosynthesis; Templates, Genetic; Parvovirus; DNA polymerase; Rodentia; Replication initiation; Virus; Vertebrata; Mammalia; Mouse; Parvoviridae; Animal; DNA; Primase; Replication; Mechanism of action; Mouse minute virus
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.82.12.4023

The distribution of termination and initiation sites in a 5081-nucleotide minute virus of mice DNA template being copied by a highly purified mouse DNA polymerase α -DNA primase complex in the presence of GTP has been examined. The 3′-hydroxyl termini (17 in all) were clustered at six sites that were located 2-14 nucleotides upstream of C2A2C2, C2AC3, or C2A2T2 sequences. When either [α -32P]- or [γ -32P]GTP was included in the DNA polymerase reaction mixtures, nascent DNA became radiolabeled. Analysis of the 32P-labeled material following treatment of the DNA with tobacco acid pyrophosphatase, bacterial alkaline phosphatase, or ribonuclease T1 revealed the presence of oligoribonucleotide chains averaging 5-7 nucleotides long and beginning with 5′ GTP residues. Eight presumptive DNA primase initiation sites were located opposite C4 or C5 sequences 3-9 nucleotides upstream of one of the three closely related hexanucleotides C2A2C2, C2AC3, and C2A2T2. RNA--DNA junctions were found 3-10 nucleotides downstream of DNA primase initiation sites. The results indicate that hexanucleotides having the general formula C2A1-2(C2-3/T<l atex>$_{2}$ ), herein referred to as ψ , are involved in promoting termination of DNA synthesis and/or de novo initiation of RNA-primed DNA chains by DNA polymerase α -primase.