The POL1 gene from the fission yeast, Schizosaccharomyces pombe, shows conserved amino acid blocks specific for eukaryotic DNA polymerases alpha

Damagnez, Véronique; Tillit, Jeanne; Recondo, A M de; Baldacci, Giuseppe

doi:10.1007/bf00273602

Cited by 58 publications

(26 citation statements)

References 22 publications

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“…6 using TB19. This construct contains the N-terminal 190 amino acid residues of DNA polymerase a; TB19 contains putative NLSs (Table 3), but lacks the catalytic domain of the enzyme (Damagnez et al 1991). The intensity of nuclear¯uorescence in TB19 also showed characteristic changes during the mitotic and meiotic cell cycles.…”

Section: Live Observation Of Gfp-fusion Constructsmentioning

confidence: 99%

Large‐scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP‐fusion genomic DNA library

Ding¹,

Tomita²,

Yamamoto³

et al. 2000

Genes to Cells

140

109

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Section: Live Observation Of Gfp-fusion Constructsmentioning

confidence: 99%

Large‐scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP‐fusion genomic DNA library

Ding¹,

Tomita²,

Yamamoto³

et al. 2000

Genes to Cells

140

109

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“…Peptide sequences of bovine p68 subunit were determined as described (Eckerskorn et al, 1988;Eckerskorn and Lottspeich, 1989) and the peptide FTLETLNSFEHEFLSK was used to synthesize two reverse complement, degenerate oligonucleotides p68 4k and p68 5k [p68 4k, 5 (Wong et al, 1988;Melov et al, 1992). Conserved regions of p180 are underlined and named with letters or Roman numerals above the amino acid sequences (Damagnez et al, 1991). The primers used for DNA amplification and the 5'-3' polarity are indicated by the arrows with the names underneath them.…”

Section: Cdna Cloning Of Dna-polymerase-a-primase Subunitsmentioning

confidence: 99%

“…The cDNAs encoding the subunits of DNA-polymerasea-primase from several eukaryotes have been cloned and sequenced; cDNAs for all four subunits from yeast and mouse, p180 DNA polymerase a and p73 subunit from Drosophila, human p180 DNA polymerase a and, recently, p68 (Lucchini et al, 1987;Pizzagalli et al, 1988;Wong et al, 1988;Foiani et al, 1989a;Prussak et al, 1989a;Brooke et al, 1991;Damagnez et al, 1991;Hirose et al, 1991;Cotterill et al, 1992;Melov et al, 1992;Collins et al, 1993;Miyazawa et al, 1993). Sequence comparisons showed that p180 DNA polymerase a is distantly related to DNA polymerases from bacteriophages, viruses and bacteria and contains several regions that are highly conserved among the mammalian, Drosophila and yeast p180 subunits (Braithwaite and Ito, 1993).…”

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confidence: 99%

DNA replication in vitro by recombinant DNA‐polymerase‐α‐primase

Stadlbauer

Brueckner

Rehfuess

et al. 1994

European Journal of Biochemistry

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DNA-polymerase-a-primase complex contains four subunits, pl80, p68, p58, and p48, and comprises a minimum of two enzymic functions. We have cloned cDNAs encoding subunits of DNA-polymerase-a-primase from human and mouse. Sequence comparisons showed high amino acid conservation among the mammalian proteins. We have over-expressed the single polypeptides and co-expressed various subunit complexes using baculovirus vectors, purified the proteins and investigated their biochemical properties. The purified mouse p48 subunit (Mp48) alone had primase activity. Purification of co-expressed Mp48 and Mp58 subunits yielded stable DNA primase of high specific activity. Co-expression of all four subunits yielded large quantities of tetrameric DNApolymerase-a-primase. The pl80, p58 and p48 polypeptides were also co-expressed and immunoaffinity purified as a trimeric enzyme complex. The tetrameric and trimeric DNA-polymerase-aprimase complexes showed both DNA primase and DNA polymerase activities. The tetrameric recombinant DNA-polymerase-a-primase synthesized double-stranded M13 DNA and replicated polyoma viral DNA in vitro efficiently.The study of enzymic mechanisms, fidelity and regulation of DNA replication is fundamental to a detailed understanding of the stability of genetic information. DNA replication of eukaryotic genomes can be divided into several steps; recognition of an origin of DNA replication, assembly of a pre-initiation complex, unwinding of origin DNA, initiation of replication within an origin and elongation of leading and lagging strands (Kornberg and Baker, 1991). DNA polymerase a and DNA primase are essential components of the cellular DNA replication machinery (Pizzagalli et al., 1988 ; Francesconi et al., 1991), playing an essential role both in initiation and in lagging strand synthesis (Thommes and Hubscher, 1990;Kornberg and Baker, 1991 ;.DNA primase and DNA polymerase a co-purify as a complex containing four subunits with molecular masses of 180, 68, 58 and 48kDa (Thommes and Hubscher, 1990;. The p380 subunit of DNA-polymerase-a-primase carries the DNA polymerase activity, and is phosphorylated in a cell-cycle-dependent manner (Pizzagalli et al.,

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“…ϩ , and pcn1 ϩ , encoding the catalytic subunits of Pol ␣, Pol ␦, Pol ε, and PCNA, respectively, have also been identified (12,14,23,46,61).…”

mentioning

confidence: 99%

Fission Yeast Cdc24 Is a Replication Factor C- and Proliferating Cell Nuclear Antigen-Interacting Factor Essential for S-Phase Completion

Tanaka

Murakami

et al. 1999

Molecular and Cellular Biology

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At the nonpermissive temperature the fission yeast cdc24-M38 mutant arrests in the cell cycle with incomplete DNA replication as indicated by pulsed-field gel electrophoresis. The cdc24 ؉ gene encodes a 501-aminoacid protein with no significant homology to any known proteins. The temperature-sensitive cdc24 mutant is effectively rescued by pcn1 ؉ , rfc1 ؉ (a fission yeast homologue of RFC1), and hhp1 ؉ , which encode the proliferating cell nuclear antigen (PCNA), the large subunit of replication factor C (RFC), and a casein kinase I involved in DNA damage repair, respectively. The Cdc24 protein binds PCNA and RFC1 in vivo, and the domains essential for Cdc24 function and for RFC1 and PCNA binding colocalize in the N-terminal two-thirds of the molecule. In addition, cdc24؉ genetically interacts with the gene encoding the catalytic subunit of DNA polymerase , which is stimulated by PCNA and RFC, and with those encoding the fission yeast counterparts of Mcm2, Mcm4, and Mcm10. These results indicate that Cdc24 is an RFC-and PCNA-interacting factor required for DNA replication and might serve as a target for regulation.Chromosomal DNA is replicated by cooperation of a number of factors and enzymes. In the budding yeast Saccharomyces cerevisiae they include the origin recognition complex, which is composed of the six subunits called Orc1 to Orc6 (4); Cdc6 (8); minichromosome maintenance (MCM) proteins (56); and at least three DNA polymerases. The origin recognition complex recognizes and binds origins of replication throughout the cell cycle. When cells enter S phase, MCM proteins are loaded onto the origins in a Cdc6-dependent manner (2, 53), and finally DNA polymerase ␣ (Pol ␣), Pol ␦, and Pol ε are recruited to start DNA synthesis. A recent analysis indicates that not only these polymerases but also some MCM proteins are components of replication forks (2).The roles of Pol ␣, Pol ␦, and other factors in DNA replication were initially elucidated by studies with a cell-free simian virus 40 (SV40) DNA replication system (reviewed in references 51 and 58). In this system, Pol ␣ with its primase subunits synthesizes RNA primers and elongates them to short initiator DNAs, which in turn serve as primers for the leadingand lagging-strand synthesis that is catalyzed by Pol ␦. Thus, during DNA replication, the polymerase used for synthesis is switched from ␣ to ␦. This switch requires two accessory proteins, replication factor C (RFC) and proliferating cell nuclear antigen (PCNA) (55). RFC binds the primer ends and thereby recruits and loads PCNA onto them (29, 54). Subsequently, Pol ␦ binds the complex and elongates the DNA strands with high processivity (54). RFC is composed of five related subunits, one large and four small ones. By contrast, PCNA forms homotrimers to act as a sliding clamp (27). This factor is essential for the high processivity of Pol ␦ (47, 48). These polymerases and accessory proteins are highly conserved throughout eukaryotes. The replication of chromosomal DNA, however, requires another type of DNA ...

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The POL1 gene from the fission yeast, Schizosaccharomyces pombe, shows conserved amino acid blocks specific for eukaryotic DNA polymerases alpha

Cited by 58 publications

References 22 publications

Large‐scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP‐fusion genomic DNA library

Large‐scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP‐fusion genomic DNA library

DNA replication in vitro by recombinant DNA‐polymerase‐α‐primase

Fission Yeast Cdc24 Is a Replication Factor C- and Proliferating Cell Nuclear Antigen-Interacting Factor Essential for S-Phase Completion

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