ATP-dependent structural change of the eukaryotic clamp-loader protein, replication factor C

被引:65
作者
Shiomi, Y
Usukura, J
Masamura, Y
Takeyasu, K
Nakayama, Y
Obuse, C
Yoshikawa, H
Tsurimoto, T [1 ]
机构
[1] Nara Inst Sci & Technol, Nara 6300101, Japan
[2] Nagoya Univ, Sch Med, Dept Anat, Nagoya, Aichi 4660065, Japan
[3] Kyoto Univ, Grad Sch Biostudies, Lab Plasma Membrane & Nucl Signaling, Kyoto 6068591, Japan
[4] Univ Osaka Prefecture, Dept Phys & Elect, Osaka 5998531, Japan
关键词
D O I
10.1073/pnas.97.26.14127
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The eukaryotic DNA sliding clamp that keeps DNA polymerase engaged at a replication fork, called proliferating cell nuclear antigen (PCNA), is loaded onto the 3' ends of primer DNA through its interaction with a heteropentameric protein complex called replication factor C (RFC). The ATPase activity of RFC is necessary for formation of a functional PCNA clamp. In the present study, the sensitivity of RFC to partial proteolysis is used to show that addition of ATP, ATP gammaS, or ADP induces different structural changes in RFC. Direct observation by electron microscopy reveals that RFC has a closed two-finger structure called the U form in the absence of ATP. This is converted into a more open C form on addition of ATP. in contrast, the structural changes induced by ATP gammaS or ADP are limited. These results suggest that RFC adapts on opened configuration intermediately after ATP hydrolysis. We further observe that PCNA is held between the two fingers of RFC and propose that the RFC structure change we observe during ATP hydrolysis causes the attached PCNA to form its active ring-like clamp on DNA.
引用
收藏
页码:14127 / 14132
页数:6
相关论文
共 49 条
  • [11] Overproduction and affinity purification of Saccharomyces cerevisiae replication factor C
    Gerik, KJ
    Gary, SL
    Burgers, PMJ
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (02) : 1256 - 1262
  • [12] Structure of the C-terminal region of p21(WAF1/CIP1) complexed with human PCNA
    Gulbis, JM
    Kelman, Z
    Hurwitz, J
    ODonnell, M
    Kuriyan, J
    [J]. CELL, 1996, 87 (02) : 297 - 306
  • [13] Division of labor - sequential ATP hydrolysis drives assembly of a DNA polymerase sliding clamp around DNA
    Hingorani, MM
    Bloom, LB
    Goodman, MF
    O'Donnell, M
    [J]. EMBO JOURNAL, 1999, 18 (18) : 5131 - 5144
  • [14] ATP binding to the Escherichia coli clamp loader powers opening of the ring-shaped clamp of DNA polymerase III holoenzyme
    Hingorani, MM
    O'Donnell, M
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (38) : 24550 - 24563
  • [15] Cleavage of BP180, a 180-kDa bullous pemphigoid antigen, yields a 120-kDa collagenous extracellular polypeptide
    Hirako, Y
    Usukura, J
    Uematsu, J
    Hashimoto, T
    Kitajima, Y
    Owaribe, K
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (16) : 9711 - 9717
  • [16] Demonstration of the molecular shape of BP180, a 180-kDa bullous pemphigoid antigen and its potential for trimer formation
    Hirako, Y
    Usukura, J
    Nishizawa, Y
    Owaribe, K
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (23) : 13739 - 13745
  • [17] HUBSCHER U, 1996, DNA REPLICATION EUKA, P525
  • [18] Proliferating cell nuclear antigen: more than a clamp for DNA polymerases
    Jonsson, ZO
    Hubscher, U
    [J]. BIOESSAYS, 1997, 19 (11) : 967 - 975
  • [19] MCM proteins: evolution, properties, and role in DNA replication
    Kearsey, SE
    Labib, K
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION, 1998, 1398 (02): : 113 - 136
  • [20] PCNA: Structure, functions and interactions
    Kelman, Z
    [J]. ONCOGENE, 1997, 14 (06) : 629 - 640