The yeast DNA polymerase-primase complex: genes and proteins.
The yeast DNA polymerase-primase complex is composed of four polypeptides designated p180, p74, p58 and p48. All the genes coding for these polypeptides have now been cloned. By protein sequence comparison we found that yeast DNA polymerase I (alpha) shares three major regions of homology with several DNA polymerases. A fourth region, called region P, is conserved in yeast and human DNA polymerase alpha. The site of a temperature-sensitive mutation in the POL1 gene which causes decreased stability of the polymerase-primase complex has been sequenced and falls in this region. We hypothesize that region P is important for protein-protein interactions. Highly selective biochemical methods might be similarly important to distinguish functional domains in the polymerase-primase complex. An autocatalytic affinity labeling procedure has been applied to map the active center of yeast DNA primase. From this approach we conclude that both primase subunits (p48 and p58) participate in the formation of the catalytic site of the enzyme.
Plevani
P
P
Dipartimento di Genetica, Università degli Studi di Milano, Italy.
Foiani
M
M
Muzi Falconi
M
M
Pizzagalli
A
A
Santocanale
C
C
Francesconi
S
S
Valsasnini
P
P
Comedini
A
A
Piatti
S
S
Lucchini
G
G
D000595:Amino Acid Sequence; D001665:Binding Sites; D004256:DNA Polymerase I / Q000235:genetics; D004257:DNA Polymerase II / Q000235:genetics; D019915:DNA Primase; D004261:DNA Replication; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007118:Immunoassay; D009154:Mutation; D012316:RNA Nucleotidyltransferases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012689:Sequence Homology, Nucleic Acid; D013329:Structure-Activity Relationship
D019915:DNA Primase; D012316:RNA Nucleotidyltransferases; D004256:DNA Polymerase I; D004257:DNA Polymerase II; D004259:DNA-Directed DNA Polymerase