| Literature DB >> 27184763 |
Hector Guillen-Ahlers1, Prahlad K Rao2, Mark E Levenstein3, Julia Kennedy-Darling3, Danu S Perumalla2, Avinash Y L Jadhav2, Jeremy P Glenn2, Amy Ludwig-Kubinski4, Eugene Drigalenko2, Maria J Montoya2, Harald H Göring2, Corianna D Anderson4, Mark Scalf3, Heidi I S Gildersleeve2, Regina Cole4, Alexandra M Greene4, Akua K Oduro5, Katarina Lazarova4, Anthony J Cesnik3, Jared Barfknecht4, Lisa A Cirillo5, Audrey P Gasch6, Michael R Shortreed3, Lloyd M Smith3, Michael Olivier7.
Abstract
Currently available methods for interrogating DNA-protein interactions at individual genomic loci have significant limitations, and make it difficult to work with unmodified cells or examine single-copy regions without specific antibodies. In this study, we describe a physiological application of the Hybridization Capture of Chromatin-Associated Proteins for Proteomics (HyCCAPP) methodology we have developed. Both novel and known locus-specific DNA-protein interactions were identified at the ENO2 and GAL1 promoter regions of Saccharomyces cerevisiae, and revealed subgroups of proteins present in significantly different levels at the loci in cells grown on glucose versus galactose as the carbon source. Results were validated using chromatin immunoprecipitation. Overall, our analysis demonstrates that HyCCAPP is an effective and flexible technology that does not require specific antibodies nor prior knowledge of locally occurring DNA-protein interactions and can now be used to identify changes in protein interactions at target regions in the genome in response to physiological challenges.Entities:
Keywords: Chromatin; Chromatin immunoprecipitation; DNA-protein interactions; ENO2; Proteomics; Yeast
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Year: 2016 PMID： 27184763 PMCID： PMC5017017 DOI： 10.1016/j.ygeno.2016.05.002
Source DB: PubMed Journal: Genomics ISSN： 0888-7543 Impact factor: 5.736