• Something wrong with this record ?

The PCNA interaction protein box sequence in Rad54 is an integral part of its ATPase domain and is required for efficient DNA repair and recombination

RC. Burgess, M. Sebesta, A. Sisakova, VP. Marini, M. Lisby, J. Damborsky, H. Klein, R. Rothstein, L. Krejci,

. 2013 ; 8 (12) : e82630.

Language English Country United States

Document type Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't

Persistent link   https://www.medvik.cz/link/bmc15014556

Rad54 is an ATP-driven translocase involved in the genome maintenance pathway of homologous recombination (HR). Although its activity has been implicated in several steps of HR, its exact role(s) at each step are still not fully understood. We have identified a new interaction between Rad54 and the replicative DNA clamp, proliferating cell nuclear antigen (PCNA). This interaction was only mildly weakened by the mutation of two key hydrophobic residues in the highly-conserved PCNA interaction motif (PIP-box) of Rad54 (Rad54-AA). Intriguingly, the rad54-AA mutant cells displayed sensitivity to DNA damage and showed HR defects similar to the null mutant, despite retaining its ability to interact with HR proteins and to be recruited to HR foci in vivo. We therefore surmised that the PCNA interaction might be impaired in vivo and was unable to promote repair synthesis during HR. Indeed, the Rad54-AA mutant was defective in primer extension at the MAT locus as well as in vitro, but additional biochemical analysis revealed that this mutant also had diminished ATPase activity and an inability to promote D-loop formation. Further mutational analysis of the putative PIP-box uncovered that other phenotypically relevant mutants in this domain also resulted in a loss of ATPase activity. Therefore, we have found that although Rad54 interacts with PCNA, the PIP-box motif likely plays only a minor role in stabilizing the PCNA interaction, and rather, this conserved domain is probably an extension of the ATPase domain III.

References provided by Crossref.org

000      
00000naa a2200000 a 4500
001      
bmc15014556
003      
CZ-PrNML
005      
20150421092214.0
007      
ta
008      
150420s2013 xxu f 000 0|eng||
009      
AR
024    7_
$a 10.1371/journal.pone.0082630 $2 doi
035    __
$a (PubMed)24376557
040    __
$a ABA008 $b cze $d ABA008 $e AACR2
041    0_
$a eng
044    __
$a xxu
100    1_
$a Burgess, Rebecca C $u Department of Genetics & Development, Columbia University Medical Center, New York, New York, United States of America.
245    14
$a The PCNA interaction protein box sequence in Rad54 is an integral part of its ATPase domain and is required for efficient DNA repair and recombination / $c RC. Burgess, M. Sebesta, A. Sisakova, VP. Marini, M. Lisby, J. Damborsky, H. Klein, R. Rothstein, L. Krejci,
520    9_
$a Rad54 is an ATP-driven translocase involved in the genome maintenance pathway of homologous recombination (HR). Although its activity has been implicated in several steps of HR, its exact role(s) at each step are still not fully understood. We have identified a new interaction between Rad54 and the replicative DNA clamp, proliferating cell nuclear antigen (PCNA). This interaction was only mildly weakened by the mutation of two key hydrophobic residues in the highly-conserved PCNA interaction motif (PIP-box) of Rad54 (Rad54-AA). Intriguingly, the rad54-AA mutant cells displayed sensitivity to DNA damage and showed HR defects similar to the null mutant, despite retaining its ability to interact with HR proteins and to be recruited to HR foci in vivo. We therefore surmised that the PCNA interaction might be impaired in vivo and was unable to promote repair synthesis during HR. Indeed, the Rad54-AA mutant was defective in primer extension at the MAT locus as well as in vitro, but additional biochemical analysis revealed that this mutant also had diminished ATPase activity and an inability to promote D-loop formation. Further mutational analysis of the putative PIP-box uncovered that other phenotypically relevant mutants in this domain also resulted in a loss of ATPase activity. Therefore, we have found that although Rad54 interacts with PCNA, the PIP-box motif likely plays only a minor role in stabilizing the PCNA interaction, and rather, this conserved domain is probably an extension of the ATPase domain III.
650    _2
$a adenosintrifosfatasy $x chemie $7 D000251
650    _2
$a aminokyselinové motivy $7 D020816
650    _2
$a sekvence aminokyselin $7 D000595
650    _2
$a párování chromozomů $7 D023902
650    _2
$a konzervovaná sekvence $7 D017124
650    _2
$a DNA $x metabolismus $7 D004247
650    _2
$a poškození DNA $7 D004249
650    _2
$a DNA-helikasy $x chemie $x metabolismus $7 D004265
650    _2
$a mutační analýza DNA $7 D004252
650    _2
$a DNA primery $x metabolismus $7 D017931
650    12
$a oprava DNA $7 D004260
650    _2
$a enzymy opravy DNA $x chemie $x metabolismus $7 D045643
650    _2
$a nestabilita genomu $7 D042822
650    _2
$a molekulární sekvence - údaje $7 D008969
650    _2
$a mutantní proteiny $x metabolismus $7 D050505
650    _2
$a mutace $x genetika $7 D009154
650    _2
$a proliferační antigen buněčného jádra $x metabolismus $7 D018809
650    _2
$a vazba proteinů $7 D011485
650    _2
$a multimerizace proteinu $7 D055503
650    _2
$a terciární struktura proteinů $7 D017434
650    12
$a rekombinace genetická $7 D011995
650    _2
$a Saccharomyces cerevisiae $x genetika $x metabolismus $7 D012441
650    _2
$a Saccharomyces cerevisiae - proteiny $x chemie $x metabolismus $7 D029701
650    _2
$a vztahy mezi strukturou a aktivitou $7 D013329
655    _2
$a časopisecké články $7 D016428
655    _2
$a Research Support, N.I.H., Extramural $7 D052061
655    _2
$a práce podpořená grantem $7 D013485
700    1_
$a Sebesta, Marek $u National Centre for Biomolecular Research, Masaryk University, Brno, Czech Republic ; Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic ; International Clinical Research Centre, Centre for Biomolecular and Cellular Engineering, Saint Anne's University Hospital, Brno, Czech Republic.
700    1_
$a Sisakova, Alexandra $u Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic ; International Clinical Research Centre, Centre for Biomolecular and Cellular Engineering, Saint Anne's University Hospital, Brno, Czech Republic.
700    1_
$a Marini, Victoria P $u Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
700    1_
$a Lisby, Michael $u Department of Molecular Biology, University of Copenhagen, Copenhagen N, Denmark.
700    1_
$a Damborsky, Jiri $u International Clinical Research Centre, Centre for Biomolecular and Cellular Engineering, Saint Anne's University Hospital, Brno, Czech Republic ; Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic.
700    1_
$a Klein, Hannah $u Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, New York, United States of America.
700    1_
$a Rothstein, Rodney $u Department of Genetics & Development, Columbia University Medical Center, New York, New York, United States of America.
700    1_
$a Krejci, Lumir $u National Centre for Biomolecular Research, Masaryk University, Brno, Czech Republic ; Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic ; International Clinical Research Centre, Centre for Biomolecular and Cellular Engineering, Saint Anne's University Hospital, Brno, Czech Republic.
773    0_
$w MED00180950 $t PloS one $x 1932-6203 $g Roč. 8, č. 12 (2013), s. e82630
856    41
$u https://pubmed.ncbi.nlm.nih.gov/24376557 $y Pubmed
910    __
$a ABA008 $b sig $c sign $y a $z 0
990    __
$a 20150420 $b ABA008
991    __
$a 20150421092513 $b ABA008
999    __
$a ok $b bmc $g 1072137 $s 897434
BAS    __
$a 3
BAS    __
$a PreBMC
BMC    __
$a 2013 $b 8 $c 12 $d e82630 $i 1932-6203 $m PLoS One $n PLoS One $x MED00180950
LZP    __
$a Pubmed-20150420

Find record

Citation metrics

    Archiving options