Genome‐wide protein interaction maps using two‐hybrid systems

FEBS Letters

Legrain, Pierre¹  Selig, Luc¹ 

[1] Hybrigenics, 180 Avenue Daumesnil, Paris 75012, France

关键词: Biological network;    Pathway;    Library screening;    Protein array;    Two-hybrid;   

DOI  :  10.1016/S0014-5793(00)01774-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Automated sequence technology has rendered functional biology amenable to genomic scale analysis. Among genome-wide exploratory approaches, the two-hybrid system in yeast (Y2H) has outranked other techniques because it is the system of choice to detect protein–protein interactions. Deciphering the cascade of binding events in a whole cell helps define signal transduction and metabolic pathways or enzymatic complexes. The function of proteins is eventually attributed through whole cell protein interaction maps where totally unknown proteins are partnered with fully annotated proteins belonging to the same functional category. Since its first description in the late 1980's, several versions of the Y2H have been developed in order to overcome the major limitations of the system, namely false positives and false negatives. Optimized versions have been recently applied at multi-molecular and genomic scale. These genome-wide surveys can be methodologically divided into two types of approaches: one either tests combinations of predefined polypeptides (the so-called matrix approach) using various short-cuts to speed up the process, or one screens with a given polypeptide (bait) for potential partners (preys) present in complex libraries of genomic or complementary DNA (library screening). In the former strategy, one tests what one knows, for example pair-wise interactions between full-length open reading frames from recently sequenced and annotated genomes. Although based on a one-by-one scheme, this method is reported to be amenable to large-scale genomics thanks to multicloning strategies and to the use of small robotics workstations. In the latter, highly complex cDNA or genomic libraries of protein domains can be screened to saturation with high-throughput screening systems allowing the discovery of yet unidentified proteins. Both approaches have strengths and drawbacks that will be discussed here. None yields a full proteome-wide screening since certain proteins (e.g. some transcription factors) are not usable in Y2H. Novel two-hybrid assays have been recently described in bacteria. Applications of these time- and cost-effective assays to genomic screening will be discussed and compared to the Y2H technology.

【 授权许可】

Unknown