Torsional regulation of hRPA-induced unwinding of double-stranded DNA

被引：35

作者：

De Vlaminck, Iwijn ^{[1
]}

Vidic, Iztok ^{[2
]}

van Loenhout, Marijn T. J. ^{[1
]}

Kanaar, Roland ^{[2
,3
]}

Lebbink, Joyce H. G. ^{[2
,3
]}

Dekker, Cees ^{[1
]}

机构：

[1] Delft Univ Technol, Kavli Inst Nanosci, NL-2628 CJ Delft, Netherlands

[2] Canc Genom Ctr, Dept Cell Biol & Genet, NL-3000 CA Rotterdam, Netherlands

[3] Erasmus MC, Dept Radiat Oncol, NL-3000 CA Rotterdam, Netherlands

来源：

NUCLEIC ACIDS RESEARCH | 2010年 / 38卷 / 12期

关键词：

REPLICATION PROTEIN-A; NUCLEIC-ACIDS; BINDING; RPA; RECOGNITION; MECHANISM; ASSOCIATION; GENERATION; FILAMENTS; DYNAMICS;

D O I：

10.1093/nar/gkq067

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

All cellular single-stranded (ss) DNA is rapidly bound and stabilized by single stranded DNA-binding proteins (SSBs). Replication protein A, the main eukaryotic SSB, is able to unwind double-stranded (ds) DNA by binding and stabilizing transiently forming bubbles of ssDNA. Here, we study the dynamics of human RPA (hRPA) activity on topologically constrained dsDNA with single-molecule magnetic tweezers. We find that the hRPA unwinding rate is exponentially dependent on torsion present in the DNA. The unwinding reaction is self-limiting, ultimately removing the driving torsional stress. The process can easily be reverted: release of tension or the application of a rewinding torque leads to protein dissociation and helix rewinding. Based on the force and salt dependence of the in vitro kinetics we anticipate that the unwinding reaction occurs frequently in vivo. We propose that the hRPA unwinding reaction serves to protect and stabilize the dsDNA when it is structurally destabilized by mechanical stresses.

引用

页码：4133 / 4142

页数：10

共 42 条

[1] Structure of the major single-stranded DNA-binding domain of replication protein A suggests a dynamic mechanism for DNA binding [J].

Bochkareva, E ;

Belegu, V ;

Korolev, S ;

Bochkarev, A .

EMBO JOURNAL, 2001, 20 (03) :612-618

[2] Structure of the RPA trimerization core and its role in the multistep DNA-binding mechanism of RPA [J].

Bochkareva, E ;

Korolev, S ;

Lees-Miller, SP ;

Bochkarev, A .

EMBO JOURNAL, 2002, 21 (07) :1855-1863

[3] Structural characterization of human RPA sequential binding to single-stranded DNA using ssDNA as a molecular ruler [J].

Cai, Lifeng ;

Roginskaya, Marina ;

Qu, Youxing ;

Yang, Zhengguan ;

Xu, Ying ;

Zou, Yue .

BIOCHEMISTRY, 2007, 46 (28) :8226-8233

[4] A dynamic model for replication protein A (RPA) function in DNA processing pathways [J].

Fanning, Ellen ;

Klimovich, Vitaly ;

Nager, Andrew R. .

NUCLEIC ACIDS RESEARCH, 2006, 34 (15) :4126-4137

[5] Abrupt buckling transition observed during the plectoneme formation of individual DNA molecules [J].

Forth, Scott ;

Deufel, Christopher ;

Sheinin, Maxim Y. ;

Daniels, Bryan ;

Sethna, James P. ;

Wang, Michelle D. .

PHYSICAL REVIEW LETTERS, 2008, 100 (14)

[6] DNA UNWINDING ACTIVITY OF REPLICATION PROTEIN-A [J].

GEORGAKI, A ;

STRACK, B ;

PODUST, V ;

HUBSCHER, U .

FEBS LETTERS, 1992, 308 (03) :240-244

[7] DNA UNWINDING BY REPLICATION PROTEIN-A IS A PROPERTY OF THE 70-KDA SUBUNIT AND IS FACILITATED BY PHOSPHORYLATION OF THE 32-KDA SUBUNIT [J].

GEORGAKI, A ;

HUBSCHER, U .

NUCLEIC ACIDS RESEARCH, 1993, 21 (16) :3659-3665

[8] Generation of superhelical torsion by ATP-dependent chromatin remodeling activities [J].

Havas, K ;

Flaus, A ;

Phelan, M ;

Kingston, R ;

Wade, PA ;

Lilley, DMJ ;

Owen-Hughes, T .

CELL, 2000, 103 (07) :1133-1142

[9] RPA INVOLVEMENT IN THE DAMAGE-RECOGNITION AND INCISION STEPS OF NUCLEOTIDE EXCISION-REPAIR [J].

HE, ZG ;

HENRICKSEN, LA ;

WOLD, MS ;

INGLES, CJ .

NATURE, 1995, 374 (6522) :566-569

[10]

HENRICKSEN LA, 1994, J BIOL CHEM, V269, P11121

← 1 2 3 4 5 →