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The ribosomal protein L2 interacts with the RNA polymerase alpha subunit and acts as a transcription modulator in Escherichia coli.
Rippa, Valentina; Cirulli, Claudia; Di Palo, Benedetta; Doti, Nunzianna; Amoresano, Angela; Duilio, Angela.
Afiliação
  • Rippa V; Department of Organic Chemistry and Biochemistry, School of Biotechnological Sciences, Federico II University of Naples, Naples, Italy.
J Bacteriol ; 192(7): 1882-9, 2010 Apr.
Article em En | MEDLINE | ID: mdl-20097853
Identification of interacting proteins in stable complexes is essential to understand the mechanisms that regulate cellular processes at the molecular level. Transcription initiation in prokaryotes requires coordinated protein-protein and protein-DNA interactions that often involve one or more transcription factors in addition to RNA polymerase (RNAP) subunits. The RNAP alpha subunit (RNAPalpha) is a key regulatory element in gene transcription and functions through direct interaction with other proteins to control all stages of this process. A clear description of the RNAPalpha protein partners should greatly increase our understanding of transcription modulation. A functional proteomics approach was employed to investigate protein components that specifically interact with RNAPalpha. A tagged form of Escherichia coli RNAPalpha was used as bait to determine the molecular partners of this subunit in a whole-cell extract. Among other interacting proteins, 50S ribosomal protein L2 (RPL2) was clearly identified by mass spectrometry. The direct interaction between RNAPalpha and RPL2 was confirmed both in vivo and in vitro by performing coimmunoprecipitation and bacterial two-hybrid experiments. The functional role of this interaction was also investigated in the presence of a ribosomal promoter by using a beta-galactosidase gene reporter assay. The results clearly demonstrated that RPL2 was able to increase beta-galactosidase expression only in the presence of a specific ribosomal promoter, whereas it was inactive when it was assayed with an unrelated promoter. Interestingly, other ribosomal proteins (L1, L3, L20, and L27) did not have any effect on rRNA expression. The findings reported here strongly suggest that in addition to its role in ribosome assembly the highly conserved RPL2 protein plays a specific and direct role in regulation of transcription.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas Ribossômicas / Transcrição Gênica / RNA Polimerases Dirigidas por DNA / Regulação Bacteriana da Expressão Gênica / Proteínas de Escherichia coli / Escherichia coli Tipo de estudo: Prognostic_studies Idioma: En Ano de publicação: 2010 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas Ribossômicas / Transcrição Gênica / RNA Polimerases Dirigidas por DNA / Regulação Bacteriana da Expressão Gênica / Proteínas de Escherichia coli / Escherichia coli Tipo de estudo: Prognostic_studies Idioma: En Ano de publicação: 2010 Tipo de documento: Article