Modern tools for identification of nucleic acid-binding proteins.
Numerous biological mechanisms depend on nucleic acid--protein interactions. The first step to the understanding of these mechanisms is to identify interacting molecules. Knowing one partner, the identification of other associated molecular species can be carried out using affinity-based purification procedures. When the nucleic acid-binding protein is known, the nucleic acid can be isolated and identified by sensitive techniques such as polymerase chain reaction followed by DNA sequencing or hybridization on chips. The reverse identification procedure is less straightforward in part because interesting nucleic acid-binding proteins are generally of low abundance and there are no methods to amplify amino acid sequences. In this article, we will review the strategies that have been developed to identify nucleic acid-binding proteins. We will focus on methods permitting the identification of these proteins without a priori knowledge of protein candidates.
Hégarat
Nadia
N
INSERM, U565 Case Postale 26, 57 rue Cuvier, 75231 Paris Cedex 05, France.
François
Jean-Christophe
JC
Praseuth
Danièle
D
D000818:Animals; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D030562:Databases, Protein; D006801:Humans; D046228:Microarray Analysis; D009696:Nucleic Acids / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D004268:DNA-Binding Proteins; D009696:Nucleic Acids; D016601:RNA-Binding Proteins