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Team Publications
Homologous Recombination and Cancer
Year of publication 2014
Martin Dutertre, Sarah Lambert, Aura Carreira, Mounira Amor-Guéret, Stéphan Vagner (2014 Mar
1)
DNA damage: RNA-binding proteins protect from near and far.
Trends in biochemical sciences : 141-9 : DOI : 10.1016/j.tibs.2014.01.003
Summary
Recent work, including large-scale genetic and molecular analyses, identified RNA-binding
proteins (RBPs) as major players in the prevention of genome instability. These studies show
that RBPs prevent harmful RNA/DNA hybrids and are involved in the DNA damage response
(DDR), from DNA repair to cell survival decisions. Indeed, specific RBPs allow the selective
regulation of DDR genes at multiple post-transcriptional levels (from pre-mRNA
splicing/polyadenylation to mRNA stability/translation) and are directly involved in DNA
repair. These multiple activities are mediated by RBP binding to mRNAs, nascent transcripts,
noncoding RNAs, and damaged DNA. Finally, because DNA damage modifies RBP localization
and binding to different RNA/DNA molecules, we propose that upon DNA damage, RBPs
coordinately regulate various aspects of both RNA and DNA metabolism.
Lucia Guidugli, Aura Carreira, Sandrine M Caputo, Asa Ehlen, Alvaro Galli, Alvaro N A Monteiro,
Susan L Neuhausen, Thomas V O Hansen, Fergus J Couch, Maaike P G Vreeswijk, (2014 Feb 5)
Functional assays for analysis of variants of uncertain significance in BRCA2.
Human mutation : 151-64 : DOI : 10.1002/humu.22478
Summary
Missense variants in the BRCA2 gene are routinely detected during clinical screening for
pathogenic mutations in patients with a family history of breast and ovarian cancer. These
subtle changes frequently remain of unknown clinical significance because of the lack of
genetic information that may help establish a direct correlation with cancer predisposition.
Therefore, alternative ways of predicting the pathogenicity of these variants are urgently
needed. Since BRCA2 is a protein involved in important cellular mechanisms such as DNA
repair, replication, and cell cycle control, functional assays have been developed that exploit
these cellular activities to explore the impact of the variants on protein function. In this
review, we summarize assays developed and currently utilized for studying missense
variants in BRCA2. We specifically depict details of each assay, including variants of
uncertain significance analyzed, and describe a validation set of (genetically) proven
pathogenic and neutral missense variants to serve as a golden standard for the validation of
each assay. Guidelines are proposed to enable implementation of laboratory-based methods
to assess the impact of the variant on cancer risk.
INSTITUT CURIE, 20 rue d’Ulm, 75248 Paris Cedex 05, France | 1
Team Publications
Homologous Recombination and Cancer
Year of publication 2013
Antony M Carr, Sarah Lambert, Aura Carreira (2013 Nov 29)
Replication stress-induced genome instability: the dark side of replication
maintenance by homologous recombination.
Journal of molecular biology : 4733-44 : DOI : 10.1016/j.jmb.2013.04.023
Summary
Homologous recombination (HR) is an evolutionary-conserved mechanism involved in a
subtle balance between genome stability and diversity. HR is a faithful DNA repair pathway
and has been largely characterized in the context of double-strand break (DSB) repair.
Recently, multiple functions for the HR machinery have been identified at arrested forks.
These are evident across different organisms and include replication fork-stabilization and
fork-restart functions. Interestingly, a DSB appears not to be a prerequisite for HR-mediated
replication maintenance. HR has the ability to rebuild a replisome at inactivated forks, but
perhaps surprisingly, the resulting replisome is liable to intrastrand and interstrand switches
leading to replication errors. Here, we review our current understanding of the replication
maintenance function of HR. The error proneness of these pathways leads us to suggest that
the origin of replication-associated genome instability should be re-evaluated.
Year of publication 2011
Aura Carreira, Stephen C Kowalczykowski (2011 Jun 28)
Two classes of BRC repeats in BRCA2 promote RAD51 nucleoprotein filament
function by distinct mechanisms.
Proceedings of the National Academy of Sciences of the United States of America : 10448-53 :
DOI : 10.1073/pnas.1106971108
Summary
The human tumor suppressor protein BRCA2 plays a key role in recombinational DNA repair.
BRCA2 recruits RAD51 to sites of DNA damage through interaction with eight conserved
motifs of approximately 35 amino acids, the BRC repeats; however, the specific function of
each repeat remains unclear. Here, we investigated the function of the individual BRC
repeats by systematically analyzing their effects on RAD51 activities. Our results reveal the
existence of two categories of BRC repeats that display unique functional characteristics.
One group, comprising BRC1, -2, -3, and -4, binds to free RAD51 with high affinity. The
second group, comprising BRC5, -6, -7, and -8, binds to free RAD51 with low affinity but
binds to the RAD51-ssDNA filament with high affinity. Each member of the first group
reduces the ATPase activity of RAD51, whereas none of the BRC repeats of the second group
affects this activity. Thus, through different mechanisms, both types of BRC repeats bind to
and stabilize the RAD51 nucleoprotein filament on ssDNA. In addition, members of the first
group limit binding of RAD51 to duplex DNA, where members of the second group do not.
Only the first group enhances DNA strand exchange by RAD51. Our results suggest that the
INSTITUT CURIE, 20 rue d’Ulm, 75248 Paris Cedex 05, France | 2
Team Publications
Homologous Recombination and Cancer
two groups of BRC repeats have differentially evolved to ensure efficient formation of a
nascent RAD51 filament on ssDNA by promoting its nucleation and growth, respectively. We
propose that the BRC repeats cooperate in a partially redundant but reinforcing manner to
ensure a high probability of RAD51 filament formation.
INSTITUT CURIE, 20 rue d’Ulm, 75248 Paris Cedex 05, France | 3