scholarly journals DNA polymerase α-primase complex from the silk glands of the non-mulberry silkworm Philosamia ricini

1994 ◽  
Vol 298 (3) ◽  
pp. 529-535
Author(s):  
S Niranjanakumari ◽  
K P Gopinathan
Keyword(s):  
Dna Polymerase ◽  
Larval Instar ◽  
Polymerase Activity ◽  
Dna Polymerase Delta ◽  
Philosamia Ricini ◽  
Dna Polymerase Alpha ◽  
Silk Glands ◽  
Tight Association ◽  
Mulberry Silkworm ◽  
Primase Complex

The DNA content in the silk glands of the non-mulberry silkworm Philosamia ricini increases continuously during the fourth and fifth instars of larval development indicating high levels of DNA replication in this terminally differentiated tissue. Concomitantly, the DNA polymerase alpha activity also increases in the middle and the posterior silk glands during development, reaching maximal levels in the middle of the fifth larval instar. A comparable level of DNA polymerase delta/epsilon was also observed in this highly replicative tissue. The DNA polymerase alpha-primase complex from the silk glands of P. ricini has been purified to homogeneity by conventional column chromatography as well as by immunoaffinity techniques. The molecular mass of the native enzyme is 560 kDa and the enzyme comprises six non-identical subunits. The identity of the enzyme as DNA polymerase alpha has been established by its sensitivity to inhibitors such as aphidicolin, N-ethylmaleimide, butylphenyl-dGTP, butylanilino-dATP and antibodies to polymerase alpha. The enzyme possesses primase activity capable of initiating DNA synthesis on single-stranded DNA templates. The tight association of polymerase and primase activities at a constant ratio of 6:1 is observed through all the purification steps. The 180 kDa subunit harbours the polymerase activity, while the primase activity is associated with the 45 kDa subunit.

Replication factor A is required in vivo for DNA replication, repair, and recombination

1994 ◽  
Vol 14 (12) ◽  
pp. 7884-7890
Author(s):  
M P Longhese ◽  
P Plevani ◽  
G Lucchini
Keyword(s):  
Dna Replication ◽  
Dna Polymerase ◽  
Dna Binding Protein ◽  
Dna Polymerase Delta ◽  
Gene Encoding ◽  
Replication Factor ◽  
Dna Polymerase Alpha ◽  
Essential Step ◽  
Primase Complex

Replication factor A (RF-A) is a heterotrimeric single-stranded-DNA-binding protein which is conserved in all eukaryotes. Since the availability of conditional mutants is an essential step to define functions and interactions of RF-A in vivo, we have produced and characterized mutations in the RFA1 gene, encoding the p70 subunit of the complex in Saccharomyces cerevisiae. This analysis provides the first in vivo evidence that RF-A function is critical not only for DNA replication but also for efficient DNA repair and recombination. Moreover, genetic evidence indicate that p70 interacts both with the DNA polymerase alpha-primase complex and with DNA polymerase delta.

scholarly journals Replication factor A is required in vivo for DNA replication, repair, and recombination.

1994 ◽  
Vol 14 (12) ◽  
pp. 7884-7890 ◽  
Author(s):  
M P Longhese ◽  
P Plevani ◽  
G Lucchini
Keyword(s):  
Dna Replication ◽  
Dna Polymerase ◽  
Dna Binding Protein ◽  
Dna Polymerase Delta ◽  
Gene Encoding ◽  
Replication Factor ◽  
Dna Polymerase Alpha ◽  
Essential Step ◽  
Primase Complex

Replication factor A (RF-A) is a heterotrimeric single-stranded-DNA-binding protein which is conserved in all eukaryotes. Since the availability of conditional mutants is an essential step to define functions and interactions of RF-A in vivo, we have produced and characterized mutations in the RFA1 gene, encoding the p70 subunit of the complex in Saccharomyces cerevisiae. This analysis provides the first in vivo evidence that RF-A function is critical not only for DNA replication but also for efficient DNA repair and recombination. Moreover, genetic evidence indicate that p70 interacts both with the DNA polymerase alpha-primase complex and with DNA polymerase delta.

scholarly journals Tight association of DNA primase with a subspecies of mouse DNA polymerase alpha.

1983 ◽  
Vol 258 (11) ◽  
pp. 6698-6700 ◽  
Author(s):  
T Yagura ◽  
S Tanaka ◽  
T Kozu ◽  
T Seno ◽  
D Korn
Keyword(s):  
Dna Polymerase ◽  
Dna Polymerase Alpha ◽  
Dna Primase ◽  
Tight Association

scholarly journals Subunits of the DNA polymerase alpha‐primase complex promote Notch‐mediated proliferation with discrete and shared functions in C. elegans germline

FEBS Journal ◽  
2018 ◽  
Vol 285 (14) ◽  
pp. 2590-2604 ◽  
Author(s):  
Dong Suk Yoon ◽  
Dong Seok Cha ◽  
Mohammad A. Alfhili ◽  
Brett D. Keiper ◽  
Myon‐Hee Lee
Keyword(s):  
Dna Polymerase ◽  
Dna Polymerase Alpha ◽  
C Elegans ◽  
Primase Complex

Involvement of proliferating cell nuclear antigen (cyclin) in DNA replication in living cells

1989 ◽  
Vol 9 (1) ◽  
pp. 57-66
Author(s):  
M Zuber ◽  
E M Tan ◽  
M Ryoji
Keyword(s):  
Dna Polymerase ◽  
Proliferating Cell Nuclear Antigen ◽  
Living Cells ◽  
Nuclear Antigen ◽  
Plasmid Replication ◽  
Dna Polymerase Delta ◽  
Dna Polymerase Alpha ◽  
Proliferating Cell

Proliferating cell nuclear antigen (PCNA) (also called cyclin) is known to stimulate the activity of DNA polymerase delta but not the other DNA polymerases in vitro. We injected a human autoimmune antibody against PCNA into unfertilized eggs of Xenopus laevis and examined the effects of this antibody on the replication of injected plasmid DNA as well as egg chromosomes. The anti-PCNA antibody inhibited plasmid replication by up to 67%, demonstrating that PCNA is involved in plasmid replication in living cells. This result further implies that DNA polymerase delta is necessary for plasmid replication in vivo. Anti-PCNA antibody alone did not block plasmid replication completely, but the residual replication was abolished by coinjection of a monoclonal antibody against DNA polymerase alpha. Anti-DNA polymerase alpha alone inhibited plasmid replication by 63%. Thus, DNA polymerase alpha is also required for plasmid replication in this system. In similar studies on the replication of egg chromosomes, the inhibition by anti-PCNA antibody was only 30%, while anti-DNA polymerase alpha antibody blocked 73% of replication. We concluded that the replication machineries of chromosomes and plasmid differ in their relative content of DNA polymerase delta. In addition, we obtained evidence through the use of phenylbutyl deoxyguanosine, an inhibitor of DNA polymerase alpha, that the structure of DNA polymerase alpha holoenzyme for chromosome replication is significantly different from that for plasmid replication.

scholarly journals Eukaryotic DNA primase appears to act as oligomer in DNA-polymerase-alpha-primase complex

1992 ◽  
Vol 206 (1) ◽  
pp. 7-13 ◽  
Author(s):  
Vladimir N. PODUST ◽  
Olga V. VLADIMIROVA ◽  
Elena N. MANAKOVA ◽  
Olga I. LAVRIK
Keyword(s):  
Dna Polymerase ◽  
Dna Polymerase Alpha ◽  
Dna Primase ◽  
Eukaryotic Dna ◽  
Primase Complex
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