DNA Replication Machinery:  Functional Characterization of a Complex Containing DNA Polymerase α, DNA Polymerase δ, and Replication Factor C Suggests an Asymmetric DNA Polymerase Dimer†

Biochemistry ◽  
1996 ◽  
Vol 35 (18) ◽  
pp. 5764-5777 ◽  
Author(s):  
Giovanni Maga ◽  
Ulrich Hübscher
Keyword(s):  
Dna Replication ◽  
Dna Polymerase ◽  
Functional Characterization ◽  
Replication Factor C ◽  
Replication Machinery ◽  
Replication Factor ◽  
Dna Polymerase Δ ◽  
Dna Polymerase Α ◽  
Factor C

scholarly journals Plant DNA polymerases alpha and delta mediate replication of geminiviruses

2020 ◽  
Author(s):  
Mengshi Wu ◽  
Hua Wei ◽  
Huang Tan ◽  
Shaojun Pan ◽  
Qi Liu ◽  
...  
Keyword(s):  
Dna Replication ◽  
Viral Replication ◽  
Dna Polymerase ◽  
Plant Cell ◽  
Dna Polymerases ◽  
Infected Plant ◽  
Replication Machinery ◽  
Dna Polymerase Δ ◽  
Dna Polymerase Α ◽  
Productive Replication

ABSTRACTGeminiviruses are causal agents of devastating diseases in crops. Geminiviruses have circular single-stranded (ss)DNA genomes that are replicated in the nucleus of the infected plant cell through double-stranded (ds)DNA intermediates by the plant’s DNA replication machinery; which host DNA polymerase mediates geminiviral multiplication, however, has so far remained elusive. Here, we show that subunits of the nuclear replicative DNA polymerases α and δ physically interact with the geminivirus-encoded replication enhancer protein, C3, and are required for viral replication. Our results suggest that while DNA polymerase α is essential to generate the viral dsDNA intermediate, DNA polymerase δ mediates the synthesis of new copies of the geminiviral ssDNA genome, and that the virus-encoded C3 acts selectively recruiting DNA polymerase δ over ε to favour a productive replication.

Functional sites of human PCNA which interact with p21 (Cip1/Waf1), DNA polymerase δ and replication factor C

1998 ◽  
Vol 3 (6) ◽  
pp. 357-369 ◽  
Author(s):  
Takashi Oku ◽  
Soichiro Ikeda ◽  
Hisashi Sasaki ◽  
Kotaro Fukuda ◽  
Hiroshi Morioka ◽  
...  
Keyword(s):  
Dna Polymerase ◽  
Replication Factor C ◽  
Functional Sites ◽  
Replication Factor ◽  
Dna Polymerase Δ ◽  
Factor C

DNA polymerase switching: I. Replication factor C displaces DNA polymerase α prior to PCNA loading

2000 ◽  
Vol 295 (4) ◽  
pp. 791-801 ◽  
Author(s):  
Giovanni Maga ◽  
Manuel Stucki ◽  
Silvio Spadari ◽  
Ulrich Hübscher
Keyword(s):  
Dna Polymerase ◽  
Replication Factor C ◽  
Replication Factor ◽  
Dna Polymerase Α ◽  
Factor C

scholarly journals Specific DNA replication mutations affect telomere length in Saccharomyces cerevisiae.

1996 ◽  
Vol 16 (9) ◽  
pp. 4614-4620 ◽  
Author(s):  
A K Adams ◽  
C Holm
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Replication ◽  
Telomere Length ◽  
Dna Polymerase ◽  
Structural Gene ◽  
Large Subunit ◽  
Replication Factor C ◽  
Telomere Elongation ◽  
Replication Factor ◽  
Factor C

To investigate the relationship between the DNA replication apparatus and the control of telomere length, we examined the effects of several DNA replication mutations on telomere length in Saccharomyces cerevisiae. We report that a mutation in the structural gene for the large subunit of DNA replication factor C (cdc44/rfc1) causes striking increases in telomere length. A similar effect is seen with mutations in only one other DNA replication gene: the structural gene for DNA polymerase alpha (cdc17/pol1) (M.J. Carson and L. Hartwell, Cell 42:249-257, 1985). For both genes, the telomere elongation phenotype is allele specific and appears to correlate with the penetrance of the mutations. Furthermore, fluorescence-activated cell sorter analysis reveals that those alleles that cause elongation also exhibit a slowing of DNA replication. To determine whether elongation is mediated by telomerase or by slippage of the DNA polymerase, we created cdc17-1 mutants carrying deletions of the gene encoding the RNA component of telomerase (TLC1). cdc17-1 strains that would normally undergo telomere elongation failed to do so in the absence of telomerase activity. This result implies that telomere elongation in cdc17-1 mutants is mediated by the action of telomerase. Since DNA replication involves transfer of the nascent strand from polymerase alpha to replication factor C (T. Tsurimoto and B. Stillman, J. Biol. Chem. 266:1950-1960, 1991; T. Tsurimoto and B. Stillman, J. Biol. Chem. 266:1961-1968, 1991; S. Waga and B. Stillman, Nature [London] 369:207-212, 1994), one possibility is that this step affects the regulation of telomere length.

scholarly journals The Large Subunit of Replication Factor C (Rfclp/Cdc44p) Is Required for DNA Replication and DNA Repair in Saccharomyces cerevisiae

Genetics ◽  
1996 ◽  
Vol 142 (1) ◽  
pp. 65-78 ◽  
Author(s):  
Michael A McAlear ◽  
K Michelle Tuffo ◽  
Connie Holm
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Repair ◽  
Dna Replication ◽  
Uv Radiation ◽  
Large Subunit ◽  
Restrictive Temperature ◽  
Replication Factor C ◽  
Replication Factor ◽  
Factor C

We used genetic and biochemical techniques to characterize the phenotypes associated with mutations affecting the large subunit of replication factor C (Cdc44p or Rfc1p) in Saccharomyces cerevisiae. We demonstrate that Cdc44p is required for both DNA replication and DNA repair in vivo. Cold-sensitive cdc44 mutants experience a delay in traversing S phase at the restrictive temperature following alpha factor arrest; although mutant cells eventually accumulate with a G2/M DNA content, they undergo a cell cycle arrest and initiate neither mitosis nor a new round of DNA synthesis. cdc44 mutants also exhibit an elevated level of spontaneous mutation, and they are sensitive both to the DNA damaging agent methylmethane sulfonate and to exposure to UV radiation. After exposure to UV radiation, cdc44 mutants at the restrictive temperature contain higher levels of single-stranded DNA breaks than do wild-type cells. This observation is consistent with the hypothesis that Cdc44p is involved in repairing gaps in the DNA after the excision of damaged bases. Thus, Cdc44p plays an important role in both DNA replication and DNA repair in vivo.

scholarly journals The TREX2 3′→ 5′ Exonuclease Physically Interacts with DNA Polymerase δ and Increases Its Accuracy

2002 ◽  
Vol 2 ◽  
pp. 275-281 ◽  
Author(s):  
Igor V. Shevelev ◽  
Kristijan Ramadan ◽  
Ulrich Hubscher
Keyword(s):  
Dna Replication ◽  
Dna Polymerase ◽  
Error Rates ◽  
High Fidelity ◽  
Replication Machinery ◽  
Dna Polymerase Δ ◽  
Pol I ◽  
Calf Thymus ◽  
Mutation Assay ◽  
Forward Mutation

Proofreading function by the 3′→ 5′ exonuclease of DNA polymerase δ (pol δ) is consistent with the observation that deficiency of the associated exonuclease can lead to a strong mutation phenotype, high error rates during DNA replication, and ultimately cancer. We have isolated pol δdfrom isotonic (pol δi) and detergent (pol δd) calf thymus extracts. Pol δdhad a 20-fold higher ratio of exonuclease to DNA polymerase than pol δi. This was due to the physical association of the TREX2 exonuclease to pol δd, which was missing from pol δi. Pol δdwas fivefold more accurate than pol δiunder error-prone conditions (1 μM dGTP and 20 dATP, dCTP, and dTTP) in a M13mp2 DNA forward mutation assay, and fourfold more accurate in an M13mp2T90 reversion assay. Under error-free conditions (20 μM each of the four dNTPs), however, both polymerases showed equal fidelity. Our data suggested that autonomous 3′→ 5′ exonucleases, such as TREX2, through its association with pol I can guarantee high fidelity under difficult conditions in the cell (e.g., imbalance of dNTPs) and can add to the accuracy of the DNA replication machinery, thus preventing mutagenesis.

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