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Identification of chromatin‐related protein interactions using protein microarrays
Author(s) -
Coleman Matthew A.,
Miller Kristi A.,
Beernink Peter T.,
Yoshikawa Daniel M.,
Albala Joanna S.
Publication year - 2003
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200300593
Subject(s) - chromatin , chip on chip , histone , nucleosome , dna repair , biology , non histone protein , chip sequencing , microbiology and biotechnology , high mobility group , replication protein a , histone code , hmg box , dna , genetics , dna binding protein , transcription factor , gene
Abstract Dynamic structural changes in chromatin are mediated by protein interactions that modulate multiple cellular processes including replication, transcription, recombination and DNA repair. Complexes that recognize chromatin are defined by several distinct groups of proteins that either directly modify histones or interact with histone‐DNA complexes . A protein microarray format was used to analyze the interaction of various DNA repair proteins with chromatin components. We applied proteins, antibodies and DNA to functionalized glass slides and interrogated the slides with our proteins of interest to identify novel protein‐protein interactions for proteins involved in DNA double‐strand break repair. Here we demonstrate that the DNA repair protein RAD51B, and not its cognate partner RAD51C, interacts with histones and not nucleosomes. Nucleosome‐specific interactions were demonstrated with the recently identified SWI/SNF protein, SMARCAL1. Unique RAD51B‐histone interactions were corroborated using Far Western analysis. This is the first demonstration of an interaction between RAD51B and histone proteins that may be important for the successful repair of DNA double‐strand breaks.