Characterization of a stable, major DNA polymerase alpha species devoid of DNA primase activity.
We have purified from Xenopus laevis ovaries a major DNA polymerase alpha species that lacked DNA primase activity. This primase-devoid DNA polymerase alpha species exhibited the same sensitivity as the DNA polymerase DNA primase alpha to BuAdATP and BuPdGTP, nucleotide analogs capable of distinguishing between DNA polymerase delta and DNA polymerase DNA primase alpha. The primase-devoid DNA polymerase alpha species also lacked significant nuclease activity indicative of the alpha-like (rather than delta-like) nature of the DNA polymerase. Using a poly(dT) template, the primase-devoid DNA polymerase alpha species elongated an oligo(rA10) primer up to 51-fold more effectively than an oligo(dA10) primer. In direct contrast, the DNA polymerase DNA primase alpha complex showed only a 4.6-fold preference for oligoribonucleotide primers at the same template/primer ratio. The catalytic differences between the two DNA polymerase alpha species were most dramatic at a template/primer ratio of 300. The primase-devoid DNA polymerase alpha species was found at high levels throughout oocyte and embryonic development. This suggests that the primase-devoid DNA polymerase alpha species could play a physiological role during DNA chain elongation in vivo, even if it is chemically related to DNA polymerase DNA primase alpha.
Department of Zoology, Iowa State University, Ames 50011-3223.
H B
HB
Kaiserman
R M
RM
Benbow
D000818:Animals; D002850:Chromatography, Gel; D004257:DNA Polymerase II / Q000302:isolation & purification / Q000378:metabolism*; D019915:DNA Primase; D004261:DNA Replication; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D008970:Molecular Weight; D010053:Ovary / Q000201:enzymology; D012316:RNA Nucleotidyltransferases / Q000378:metabolism*; D013379:Substrate Specificity; D013698:Templates, Genetic; D014982:Xenopus laevis / Q000502:physiology*
D019915:DNA Primase; D012316:RNA Nucleotidyltransferases; D004257:DNA Polymerase II