A structure-function analysis of the yeast Elg1 protein reveals
the importance of PCNA unloading in genome stability
maintenance

J 2017

A structure-function analysis of the yeast Elg1 protein reveals the importance of PCNA unloading in genome stability maintenance

SHEMESH, Keren, Marek ŠEBESTA, Martin PAČESA, Soumitra SAU, Alex BRONSTEIN et. al.

Basic information

Original name

A structure-function analysis of the yeast Elg1 protein reveals the importance of PCNA unloading in genome stability maintenance

Authors

SHEMESH, Keren (376 Israel), Marek ŠEBESTA (703 Slovakia, belonging to the institution), Martin PAČESA (703 Slovakia, belonging to the institution), Soumitra SAU (376 Israel), Alex BRONSTEIN (376 Israel), Oren PARNAS (376 Israel), Batia LIEFSHITZ (376 Israel), Česlovas VENCLOVAS (440 Lithuania), Lumír KREJČÍ (203 Czech Republic, guarantor, belonging to the institution) and Martin KUPIEC (376 Israel)

Edition

Nucleic Acids Research, Oxford, Oxford University Press, 2017, 0305-1048

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

Impact factor

Impact factor: 11.561

RIV identification code

RIV/00216224:14110/17:00094734

Organization unit

Faculty of Medicine

DOI

http://dx.doi.org/10.1093/nar/gkw1348

UT WoS

000398376200030

Keywords in English

REPLICATION FACTOR-C; SISTER-CHROMATID COHESION; ALTERNATIVE RFC COMPLEX; DNA-POLYMERASE DELTA; CELL NUCLEAR ANTIGEN; SUMO-MODIFIED PCNA; SACCHAROMYCES-CEREVISIAE; ESCHERICHIA-COLI; REPAIR SYNTHESIS; CLAMP LOADERS

Abstract

V originále

The sliding clamp, PCNA, plays a central role in DNA replication and repair. In the moving replication fork, PCNA is present at the leading strand and at each of the Okazaki fragments that are formed on the lagging strand. PCNA enhances the processivity of the replicative polymerases and provides a landing platform for other proteins and enzymes. The loading of the clamp onto DNA is performed by the Replication Factor C (RFC) complex, whereas its unloading can be carried out by an RFC-like complex containing Elg1. Mutations in ELG1 lead to DNA damage sensitivity and genome instability. To characterize the role of Elg1 in maintaining genomic integrity, we used homology modeling to generate a number of site-specific mutations in ELG1 that exhibit different PCNA unloading capabilities. We show that the sensitivity to DNA damaging agents and hyper-recombination of these alleles correlate with their ability to unload PCNA from the chromatin. Our results indicate that retention of modified and unmodified PCNA on the chromatin causes genomic instability. We also show, using purified proteins, that the Elg1 complex inhibits DNA synthesis by unloading SUMOylated PCNA from the DNA. Additionally, we find that mutations in ELG1 suppress the sensitivity of rad5 Delta mutants to DNA damage by allowing translesion synthesis to take place. Taken together, the data indicate that the Elg1-RLC complex plays an important role in the maintenance of genomic stability by unloading PCNA from the chromatin.

Links

GAP207/12/2323, research and development project

Name: Endonuleazová a translokázová aktivita v restričních-modifikáčních komplexéch typu I

Investor: Czech Science Foundation

GA13-26629S, research and development project

Name: SUMO a stability genomu

Investor: Czech Science Foundation

Citovat

SHEMESH, Keren, Marek ŠEBESTA, Martin PAČESA, Soumitra SAU, Alex BRONSTEIN, Oren PARNAS, Batia LIEFSHITZ, Česlovas VENCLOVAS, Lumír KREJČÍ and Martin KUPIEC. A structure-function analysis of the yeast Elg1 protein reveals the importance of PCNA unloading in genome stability maintenance. Nucleic Acids Research. Oxford: Oxford University Press, 2017, vol. 45, No 6, p. 3189-3203. ISSN 0305-1048. Available from: https://dx.doi.org/10.1093/nar/gkw1348.

@article{1379549,
   author = {Shemesh, Keren and Šebesta, Marek and Pačesa, Martin and Sau, Soumitra and Bronstein, Alex and Parnas, Oren and Liefshitz, Batia and Venclovas, Česlovas and Krejčí, Lumír and Kupiec, Martin},
   article_location = {Oxford},
   article_number = {6},
   doi = {http://dx.doi.org/10.1093/nar/gkw1348},
   keywords = {REPLICATION FACTOR-C; SISTER-CHROMATID COHESION; ALTERNATIVE RFC COMPLEX; DNA-POLYMERASE DELTA; CELL NUCLEAR ANTIGEN; SUMO-MODIFIED PCNA; SACCHAROMYCES-CEREVISIAE; ESCHERICHIA-COLI; REPAIR SYNTHESIS; CLAMP LOADERS},
   language = {eng},
   issn = {0305-1048},
   journal = {Nucleic Acids Research},
   title = {A structure-function analysis of the yeast Elg1 protein reveals the importance of PCNA unloading in genome stability maintenance},
   volume = {45},
   year = {2017}
}

TY  - JOUR
ID  - 1379549
AU  - Shemesh, Keren - Šebesta, Marek - Pačesa, Martin - Sau, Soumitra - Bronstein, Alex - Parnas, Oren - Liefshitz, Batia - Venclovas, Česlovas - Krejčí, Lumír - Kupiec, Martin
PY  - 2017
TI  - A structure-function analysis of the yeast Elg1 protein reveals the importance of PCNA unloading in genome stability maintenance
JF  - Nucleic Acids Research
VL  - 45
IS  - 6
SP  - 3189-3203
EP  - 3189-3203
PB  - Oxford University Press
SN  - 03051048
KW  - REPLICATION FACTOR-C
KW  - SISTER-CHROMATID COHESION
KW  - ALTERNATIVE RFC COMPLEX
KW  - DNA-POLYMERASE DELTA
KW  - CELL NUCLEAR ANTIGEN
KW  - SUMO-MODIFIED PCNA
KW  - SACCHAROMYCES-CEREVISIAE
KW  - ESCHERICHIA-COLI
KW  - REPAIR SYNTHESIS
KW  - CLAMP LOADERS
N2  - The sliding clamp, PCNA, plays a central role in DNA replication and repair. In the moving replication fork, PCNA is present at the leading strand and at each of the Okazaki fragments that are formed on the lagging strand. PCNA enhances the processivity of the replicative polymerases and provides a landing platform for other proteins and enzymes. The loading of the clamp onto DNA is performed by the Replication Factor C (RFC) complex, whereas its unloading can be carried out by an RFC-like complex containing Elg1. Mutations in ELG1 lead to DNA damage sensitivity and genome instability. To characterize the role of Elg1 in maintaining genomic integrity, we used homology modeling to generate a number of site-specific mutations in ELG1 that exhibit different PCNA unloading capabilities. We show that the sensitivity to DNA damaging agents and hyper-recombination of these alleles correlate with their ability to unload PCNA from the chromatin. Our results indicate that retention of modified and unmodified PCNA on the chromatin causes genomic instability. We also show, using purified proteins, that the Elg1 complex inhibits DNA synthesis by unloading SUMOylated PCNA from the DNA. Additionally, we find that mutations in ELG1 suppress the sensitivity of rad5 Delta mutants to DNA damage by allowing translesion synthesis to take place. Taken together, the data indicate that the Elg1-RLC complex plays an important role in the maintenance of genomic stability by unloading PCNA from the chromatin.
ER  -

SHEMESH, Keren, Marek ŠEBESTA, Martin PAČESA, Soumitra SAU, Alex BRONSTEIN, Oren PARNAS, Batia LIEFSHITZ, Česlovas VENCLOVAS, Lumír KREJČÍ and Martin KUPIEC. A structure-function analysis of the yeast Elg1 protein reveals the importance of PCNA unloading in genome stability maintenance. \textit{Nucleic Acids Research}. Oxford: Oxford University Press, 2017, vol.~45, No~6, p.~3189-3203. ISSN~0305-1048. Available from: https://dx.doi.org/10.1093/nar/gkw1348.

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