Cryo-EM structures reveal high-resolution mechanism of a DNA
polymerase sliding clamp loader

J 2022

Cryo-EM structures reveal high-resolution mechanism of a DNA polymerase sliding clamp loader

GAUBITZ, Christl, Xingchen LIU, Joshua PAJAK, Nicholas P STONE, Janelle A HAYES et. al.

Basic information

Original name

Cryo-EM structures reveal high-resolution mechanism of a DNA polymerase sliding clamp loader

Authors

GAUBITZ, Christl, Xingchen LIU, Joshua PAJAK, Nicholas P STONE, Janelle A HAYES, Gabriel DEMO (703 Slovakia, guarantor, belonging to the institution) and Brian A KELCH

Edition

elife, CAMBRIDGE, ELIFE SCIENCES PUBLICATIONS LTD, 2022, 2050-084X

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10608 Biochemistry and molecular biology

Country of publisher

United Kingdom of Great Britain and Northern Ireland

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 7.700

RIV identification code

RIV/00216224:14740/22:00127475

Organization unit

Central European Institute of Technology

DOI

http://dx.doi.org/10.7554/eLife.74175

UT WoS

000766980900001

Keywords in English

sliding clamp loader; AAA plus; DNA replication; S; cerevisiae

Abstract

V originále

Sliding clamps are ring-shaped protein complexes that are integral to the DNA replication machinery of all life. Sliding clamps are opened and installed onto DNA by clamp loader AAA+ ATPase complexes. However, how a clamp loader opens and closes the sliding clamp around DNA is still unknown. Here, we describe structures of the Saccharomyces cerevisiae clamp loader Replication Factor C (RFC) bound to its cognate sliding clamp Proliferating Cell Nuclear Antigen (PCNA) en route to successful loading. RFC first binds to PCNA in a dynamic, closed conformation that blocks both ATPase activity and DNA binding. RFC then opens the PCNA ring through a large-scale ‘crab-claw’ expansion of both RFC and PCNA that explains how RFC prefers initial binding of PCNA over DNA. Next, the open RFC:PCNA complex binds DNA and interrogates the primer-template junction using a surprising base-flipping mechanism. Our structures indicate that initial PCNA opening and subsequent closure around DNA do not require ATP hydrolysis, but are driven by binding energy. ATP hydrolysis, which is necessary for RFC release, is triggered by interactions with both PCNA and DNA, explaining RFC’s switch-like ATPase activity. Our work reveals how a AAA+ machine undergoes dramatic conformational changes for achieving binding preference and substrate remodeling.

Links

LL2008, research and development project

Name: Komunikace mezi transkripcí a translací (Acronym: CROSSBETT)

Investor: Ministry of Education, Youth and Sports of the CR, Crosstalk between transcription and translation

Citovat

GAUBITZ, Christl, Xingchen LIU, Joshua PAJAK, Nicholas P STONE, Janelle A HAYES, Gabriel DEMO and Brian A KELCH. Cryo-EM structures reveal high-resolution mechanism of a DNA polymerase sliding clamp loader. elife. CAMBRIDGE: ELIFE SCIENCES PUBLICATIONS LTD, 2022, vol. 11, FEB, p. nestrankovano, 29 pp. ISSN 2050-084X. Available from: https://dx.doi.org/10.7554/eLife.74175.

@article{2240104,
   author = {Gaubitz, Christl and Liu, Xingchen and Pajak, Joshua and Stone, Nicholas P and Hayes, Janelle A and Demo, Gabriel and Kelch, Brian A},
   article_location = {CAMBRIDGE},
   article_number = {FEB},
   doi = {http://dx.doi.org/10.7554/eLife.74175},
   keywords = {sliding clamp loader; AAA plus; DNA replication; S; cerevisiae},
   language = {eng},
   issn = {2050-084X},
   journal = {elife},
   title = {Cryo-EM structures reveal high-resolution mechanism of a DNA polymerase sliding clamp loader},
   url = {https://elifesciences.org/articles/74175},
   volume = {11},
   year = {2022}
}

TY  - JOUR
ID  - 2240104
AU  - Gaubitz, Christl - Liu, Xingchen - Pajak, Joshua - Stone, Nicholas P - Hayes, Janelle A - Demo, Gabriel - Kelch, Brian A
PY  - 2022
TI  - Cryo-EM structures reveal high-resolution mechanism of a DNA polymerase sliding clamp loader
JF  - elife
VL  - 11
IS  - FEB
SP  - nestrankovano
EP  - nestrankovano
PB  - ELIFE SCIENCES PUBLICATIONS LTD
SN  - 2050084X
KW  - sliding clamp loader
KW  - AAA plus
KW  - DNA replication
KW  - S
KW  - cerevisiae
UR  - https://elifesciences.org/articles/74175
N2  - Sliding clamps are ring-shaped protein complexes that are integral to the DNA replication machinery of all life. Sliding clamps are opened and installed onto DNA by clamp loader AAA+ ATPase complexes. However, how a clamp loader opens and closes the sliding clamp around DNA is still unknown. Here, we describe structures of the Saccharomyces cerevisiae clamp loader Replication Factor C (RFC) bound to its cognate sliding clamp Proliferating Cell Nuclear Antigen (PCNA) en route to successful loading. RFC first binds to PCNA in a dynamic, closed conformation that blocks both ATPase activity and DNA binding. RFC then opens the PCNA ring through a large-scale ‘crab-claw’ expansion of both RFC and PCNA that explains how RFC prefers initial binding of PCNA over DNA. Next, the open RFC:PCNA complex binds DNA and interrogates the primer-template junction using a surprising base-flipping mechanism. Our structures indicate that initial PCNA opening and subsequent closure around DNA do not require ATP hydrolysis, but are driven by binding energy. ATP hydrolysis, which is necessary for RFC release, is triggered by interactions with both PCNA and DNA, explaining RFC’s switch-like ATPase activity. Our work reveals how a AAA+ machine undergoes dramatic conformational changes for achieving binding preference and substrate remodeling.
ER  -

GAUBITZ, Christl, Xingchen LIU, Joshua PAJAK, Nicholas P STONE, Janelle A HAYES, Gabriel DEMO and Brian A KELCH. Cryo-EM structures reveal high-resolution mechanism of a DNA polymerase sliding clamp loader. \textit{elife}. CAMBRIDGE: ELIFE SCIENCES PUBLICATIONS LTD, 2022, vol.~11, FEB, p.~nestrankovano, 29 pp. ISSN~2050-084X. Available from: https://dx.doi.org/10.7554/eLife.74175.

Detailed Information on Publication Record