孔道春
真核细胞染色体DNA 复制起始的机理与调控、Checkpoint的激活机理及其信号传导、Control of cell division cycle and cancer。
个性化签名
- 姓名:孔道春
- 目前身份:
- 担任导师情况:
- 学位:
-
学术头衔:
博士生导师
- 职称:-
-
学科领域:
物理海洋学
- 研究兴趣:真核细胞染色体DNA 复制起始的机理与调控、Checkpoint的激活机理及其信号传导、Control of cell division cycle and cancer。
男,教授,北京大学生命科学学院,研究方向是真核细胞染色体DNA 复制起始的机理与调控、Checkpoint的激活机理及其信号传导、Control of cell division cycle and cancer。 1979-1983 兰州大学生物系, 学士 1986-1989 中国科学院上海生物化学研究所,硕士 1990-1995 Temple University Medical School, USA, 博士 1995-1998 Harvard University Medical School, 博士后 1998-2000 The Memorial Sloan-Kettering Cancer Center, 博士后 2000-2005 National Institutes of Health, Bethesda, USA, 研究员
-
主页访问
3169
-
关注数
0
-
成果阅读
508
-
成果数
11
孔道春, James C.Pierce+, Daochun Kong and Warren Masker*
Nucleic Acids Research, Vol. 19, No.14 3901-3905,-0001,():
-1年11月30日
The frequency of genetic deletion between directly repeated DNA sequences in bacteriophage T7 was measured as a function of the length of the direct repeat. The non-essential ligase gene (gene 1.3) of bacteriophage T7 was interrupted with pieces of synthetic DNA bracketed by direct repeats of various lengths. Deletion of these 76 bp long inserts was too low to be measured when the direct repeats were less than 6 bp long. However, the frequency of deletion of inserts with longer direct repeats increased exponentially as the length of the repeats increased from 8 to 20 bp. When inverted repeats (palindromes) were designed in the midst of the insert there was essentially no increase in deletion frequency between 10 bp direct repeats. But, the same palindromic sequences increased the deletion frequency between 5 bp direct repeats by at least two orders of magnitude. Thus, in this system homology at the endpoints is a more important determinant of deletion frequency than is the presence of palindromes between the direct repeats.
-
35浏览
-
0点赞
-
0收藏
-
0分享
-
145下载
-
0评论
-
引用
孔道春, Daochun Kong‡ and Warren Masker‡, §
THE JOURNAOLF BIOLOGICAL CHEMISTRY Vol. 268, No.11, Issue of April 15, pp. 7721-7727.1993,-0001,():
-1年11月30日
An in vitro system based upon extracts of bacteriophage T7 infected Escherichia coli was used to study genetic deletions and to examine the importance of DNA replication in the deletion process. When T7 genomes with gene 1.3 inactivated by a 43-bp insert of random sequence DNA bracketed by 11-bp direct repeats were replicated in vitro the inserts were deleted with a frequency of about deletions per genome replication. Under conditions where deletion could take place only by recombination between direct repeats on distinct DNA molecules deletion frequency was at least an order of magnitude lower. These data demonstrate the utility of the in vitro system as a means for examining deletion mechanisms and underscore the importance of DNA replication in deletions.
-
57浏览
-
0点赞
-
0收藏
-
0分享
-
127下载
-
0评论
-
引用
【期刊论文】Deletion between Direct Repeats in T7 DNA Stimulated by Double-Strand Breaks
孔道春, DAOCHUN KoNG AND WARREN MASKER , , *
JOURNAL OF BACTERIOLOGY, Oct. 1994,P. 5904-5911,-0001,():
-1年11月30日
An in Vitro System Dased on extraCtS of Escherichia coli infected with bacteriophage T7 WaS USed to study genetic deletions between directly repeated sequences.The frequency of deletion was highest under conditions in which the DNA was actively replicating.Deletion frequency increased markedly with the length of the direct repeat DOtll ln vitro and ln VlVO.W111en a’T7 gene WaS interrupted by 93 Dp 0I nonsense SeqUerice HanKed by 20-bp direct repeats,the region between the repeats was deleted in about 1 out Of every 1,600 genomes during eaCn r0Unn 0l reDIiCanon.Vel7 Slmllar values were 10Und 10r deleU0n n.eqUen frequency ln viV0 and ln viltro1’ne deletion frequency was essentially unaffected by a recA mutation in the host.When a double.strand break wras placed between the repeats·repair of this strand break wras often accompanied by the deletion of the DNA Detween the mreCt repeats, SUggesresunR tllat Dreak rejolnlng COUId contnbUte to deletion dnnng lnⅥtr0 lJNA replication.
-
55浏览
-
0点赞
-
0收藏
-
0分享
-
122下载
-
0评论
-
引用
孔道春, Daochun Kong and Charles C.Richardson"
The EMBO Journal vol.15 No.8 pp.2010-2019, 1996,-0001,():
-1年11月30日
Two proteins encoded by bacteriophage T7, the gene 2.5 single-stranded DNA binding protein and the gene 4 helicase, mediate homologous DNA strand exchange. Gene 2.5 protein stimulates homologous base pairing of two DNA molecules containing complementary singlestranded regions. The formation of a joint molecule consisting of circular, single-stranded M13 DNA, annealed to homologous linear, duplex DNA having 3'- or 5'-single-stranded termini of -100 nucleotides requires stoichiometric amounts of gene 2.5 protein. In the presence of gene 4 helicase, strand transfer proceeds at a rate of >120 nucleotides/s in a polar 5' to 3' direction with respect to the invading strand, resulting in the production of circular duplex M13 DNA. Strand transfer is coupled to the hydrolysis of a nucleoside 5'-triphosphate. The reaction is dependent on specific interactions between gene 2.5 protein and gene 4 protein.
DNA replication/, gene 2., 5 protein/, gene 4 protein/, recombination
-
63浏览
-
0点赞
-
0收藏
-
0分享
-
160下载
-
0评论
-
引用
孔道春, Daochun Kong‡, Nancy G. Nossal§, and Charles C. Richardson‡, ¶
THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 272, No.13, Issue of March 28, pp. 8380-8387, 1997,-0001,():
-1年11月30日
Bacteriophage T7 gene 2.5 single-stranded DNA-binding protein and gene 4 DNA helicase together promote pairing of two homologous DNA molecules and subsequent polar branch migration (Kong, D., and Richardson, C. C. (1996) EMBO J. 15, 2010–2019). In this report, we show that gene 2.5 protein is not required for the initiation or propagation of strand transfer once a joint molecule has been formed between the two DNA partners, a reaction that is mediated by the gene 2.5 protein alone. A mutant gene 2.5 protein, gene 2.5-D21C protein, lacking 21 amino acid residues at its C terminus, cannot physically interact with gene 4 protein. Although it does bind to single-stranded DNA and promote the formation of joint molecule via homologous base pairing, subsequent strand transfer by gene 4 helicase is inhibited by the presence of the gene 2.5-D21C protein. Bacteriophage T4 gene 32 protein likewise inhibits T7 gene 4 protein-mediated strand transfer, whereas Escherichia coli single-stranded DNA-binding protein does not. The 63-kDa gene 4 protein of phage T7 is also a DNA primase in that it catalyzes the synthesis of oligonucleotides at specific sequences during translocation on single-stranded DNA. We find that neither the rate nor extent of strand transfer is significantly affected by concurrent primer synthesis. The bacteriophage T4 gene 41 helicase has been shown to catalyze polar branch migration after the T4 gene 59 helicase assembly protein loads the helicase onto joint molecules formed by the T4 UvsX and gene 32 proteins (Salinas, F., and Kodadek, T. (1995) Cell 82, 111-119). We find that gene 32 protein alone forms joint molecules between partially single-stranded homologous DNA partners and that subsequent branch migration requires this single-stranded DNA-binding protein in addition to the gene 41 helicase and the gene 59 helicase assembly protein. Similar to the strand transfer reaction, strand displacement DNA synthesis catalyzed by T4 DNA polymerase also requires the presence of gene 32 protein in addition to the gene 41 and 59 proteins.
-
62浏览
-
0点赞
-
0收藏
-
0分享
-
136下载
-
0评论
-
引用
孔道春, DAOCHUN KONG*, JACK D. GRIFFITH†, AND CHARLES C. RICHARDSON*, ‡
Proc. Natl. Acad. Sci. USA Vol. 94, pp. 2987-2992, April 1997,-0001,():
-1年11月30日
In bacteriophage T7 the gene 2.5 singlestranded DNA-binding protein and the gene 4 helicase together promote the annealing of homologous regions of two DNA partners to form a joint molecule and subsequent strand transfer. In this reaction T7 gene 2.5 protein is essential for joint molecule formation, but is not required for T7 gene 4 protein-mediated strand transfer. T7 gene 4 helicase alone is able to mediate strand transfer, provided that a joint molecule is available. The present paper shows that, in addition, strand transfer proceeds at a normal rate even when both DNA partners contain ultraviolet-induced pyrimidine dimers (0.6 dimer per 100 nt). An insert of a relatively long (842-nt) segment of nonhomologous DNA in the single-stranded DNA partner has no effect on strand transfer, whereas its presence in the double-stranded partner prevents strand transfer. A short insert (37 nt) can be tolerated in either partner. Thus, DNA helicase is able to participate in recombinational DNA repair through its role in strand exchange, providing a pathway distinct from nucleotide excision repair.
-
38浏览
-
0点赞
-
0收藏
-
0分享
-
78下载
-
0评论
-
引用
孔道春, Daochun Kong and Charles C. Richardson‡
THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 273, No.11, Issue of March 13, pp. 6556-6564, 1998,-0001,():
-1年11月30日
The gene 2.5 single-stranded DNA (ssDNA) binding protein of bacteriophage T7 is essential for T7 DNA replication and recombination. Earlier studies have shown that the COOH-terminal 21 amino acids of the gene 2.5 protein are essential for specific protein-protein interaction with T7 DNA polymerase and T7 DNA helicase/primase. A truncated gene 2.5 protein, in which the acidic COOH-terminal 21 amino acid residues are deleted no longer supports T7 growth, forms dimers, or interacts with either T7 DNA polymerase or T7 helicase/primase in vitro. The single-stranded DNA-binding protein encoded by Escherichia coli (SSB protein) and phage T4 (gene 32 protein) also have acidic COOH-terminal domains, but neither protein can substitute for T7 gene 2.5 protein in vivo. To determine if the specificity for the protein-protein interaction involving gene 2.5 protein resides in its COOH terminus, we replaced the COOH-terminal region of the gene 2.5 protein with the COOH-terminal region from either E. coli SSB protein or T4 gene 32 protein. Both of the two chimeric proteins can substitute for T7 gene 2.5 protein to support the growth of phage T7. The two chimeric proteins, like gene 2.5 protein, form dimers and interact with T7 DNA polymerase and helicase/primase to stimulate their activities. In contrast, chimeric proteins in which the COOH terminus of T7 gene 2.5 protein replaced the COOH terminus of E. coli SSB protein or T4 gene 32 protein cannot support the growth of phage T7. We conclude that an acidic COOH terminus of the gene 2.5 protein is essential for protein-protein interaction, but it alone cannot account for the specificity of the interaction.
-
48浏览
-
0点赞
-
0收藏
-
0分享
-
58下载
-
0评论
-
引用
孔道春, Kyeong-Yeop Moon, Daochun Kong, Joon-Kyu Lee, Santanu Raychaudhuri, and Jerard Hurwitz*
PNAS October 26, 1999 vol. 96 no. 22 12367-12372,-0001,():
-1年11月30日
The origin recognition complex (ORC), first identified in Saccharomyces cerevisiae (sc), is a six-subunit protein complex that binds to DNA origins. Here, we report the identification and cloning of cDNAs encoding the six subunits of the ORC of Schizosaccharomyces pombe (sp). Sequence analyses revealed that spOrc1, 2, and 5 subunits are highly conserved compared with their counterparts from S. cerevisiae, Xenopus, Drosophila, and human. In contrast, both spOrc3 and spOrc6 subunits are poorly conserved. As reported by Chuang and Kelly [(1999) Proc. Natl. Acad. Sci. USA 96, 2656-2661], the C-terminal region of spOrc4 is also conserved whereas the N terminus uniquely contains repeats of a sequence that binds strongly to AT-rich DNA regions. Consistent with this, extraction of S. pombe chromatin with 1 M NaCl, or after DNase I treatment, yielded the six-subunit ORC, whereas extraction with 0.3 M resulted in five-subunit ORC lacking spOrc4p. The spORC can be reconstituted in vitro with all six recombinant subunits expressed in the rabbit reticulocyte system. The association of spOrc4p with the other subunits required the removal of DNA from reaction mixture by DNase I. This suggests that a strong interaction between spOrc4p and DNA can prevent the isolation of the six-subunit ORC. The unique DNA-binding properties of the spORC may contribute to our understanding of the sequence-specific recognition required for the initiation of DNA replication in S. pombe.
replication, AT-rich sequences
-
36浏览
-
0点赞
-
0收藏
-
0分享
-
69下载
-
0评论
-
引用
孔道春, DAOCHUN KONG AND MELVIN L. DEPAMPHILIS*
MOLECULAR AND CELLULAR BIOLOGY, 8095-8103. 2001 Dec. 2001, p. 8095-8103,-0001,():
-1年11月30日
The mechanism by which origin recognition complexes (ORCs) identify replication origins was investigated using purified Orc proteins from Schizosaccharomyces pombe. Orc4p alone bound tightly and specifically to several sites within S. pombe replication origins that are genetically required for origin activity. These sites consisted of clusters of A or T residues on one strand but were devoid of either alternating A and T residues or GC-rich sequences. Addition of a complex consisting of Orc1, -2, -3, -5, and -6 proteins (ORC-5) altered neither Orc4p binding to origin DNA nor Orc4p protection of specific sequences. ORC-5 alone bound weakly and nonspecifically to DNA; strong binding required the presence of Orc4p. Under these conditions, all six subunits remained bound to chromatin isolated from each phase of the cell division cycle. These results reveal that the S. pombe ORC binds to multiple, specific sites within replication origins and that site selection, at least in vitro, is determined solely by the Orc4p subunit.
-
44浏览
-
0点赞
-
0收藏
-
0分享
-
73下载
-
0评论
-
引用
孔道春, Daochun Kong and Melvin L. DePamphilis
The EMBO Journal Vol. 21 No.20 pp. 5567-5576, 2002,-0001,():
-1年11月30日
Previous studies have shown that the Schizosaccharomyces pombe Orc4 subunit is solely responsible for in vitro binding of origin recognition complex (ORC) to speci
ARS3001/, cell cycle/, DNA replication origin/, origin recognition complex/, replication initiation point
-
29浏览
-
0点赞
-
0收藏
-
0分享
-
119下载
-
0评论
-
引用