Identification and mapping of self-assembling protein domains encoded by the Escherichia coli K-12 genome by use of lambda repressor fusions.

Abstract

Self-assembling proteins and protein fragments encoded by the Escherichia coli genome were identified from E. coli K-12 strain MG1655. Libraries of random DNA fragments cloned into a series of lambda repressor fusion vectors were subjected to selection for immunity to infection by phage lambda. Survivors were identified by sequencing the ends of the inserts, and the fused protein sequence was inferred from the known genomic sequence. Four hundred sixty-three nonredundant open reading frame-encoded interacting sequence tags (ISTs) were recovered from sequencing 2,089 candidates. These ISTs, which range from 16 to 794 amino acids in length, were clustered into families of overlapping fragments, identifying potential homotypic interactions encoded by 232 E. coli genes. Repressor fusions identified ISTs from genes in every protein-based functional category, but membrane proteins were underrepresented. The IST-containing genes were enriched for regulatory proteins and for proteins that form higher-order oligomers. Forty-eight (20.7%) homotypic proteins identified by ISTs are predicted to contain coiled coils. Although most of the IST-containing genes are identifiably related to proteins in other bacterial genomes, more than half of the ISTs do not have identifiable homologs in the Protein Data Bank, suggesting that they may include many novel structures. The data are available online at http://oligomers.tamu.edu/.

• Authors: Leonardo Mariño-Ramírez, Jonathan L Minor, Nicola Reading, James C Hu
• Journal: Journal of bacteriology (J Bacteriol) Vol. 186 Issue 5 Pg. 1311-9 (Mar 2004) ISSN: 0021-9193 [Print] United States
• PMID: 14973045 (Publication Type: Journal Article, Research Support, U.S. Gov't, Non-P.H.S., Research Support, U.S. Gov't, P.H.S.)

Chemical References

• DNA-Binding Proteins
• Escherichia coli Proteins
• Repressor Proteins
• Viral Proteins
• Viral Regulatory and Accessory Proteins
• phage repressor proteins

Topics

• Artificial Gene Fusion
• Bacteriophage lambda (genetics, metabolism)
• DNA-Binding Proteins
• Escherichia coli (chemistry, genetics, growth & development, metabolism)
• Escherichia coli Proteins (chemistry, genetics, metabolism)
• Genome, Bacterial
• Genomic Library
• Protein Interaction Mapping
• Protein Structure, Tertiary
• Repressor Proteins (genetics, metabolism)
• Sequence Tagged Sites
• Viral Proteins
• Viral Regulatory and Accessory Proteins