The RNA degradosome is a bacterial protein machine devoted to RNA turnover. Degradosomes and related complexes have been described in Escherichia coli and other prokaryotes as well as in eukaryotes. The integral components of the E. coli RNA degradosome include the endoribonuclease RNase E, the phosphorolytic exoribonuclease polynucleotide phosphorylase (PNPase), the DEAD-box helicase RhlB, and enolase, a glycolytic enzyme commonly implicated in an apparently unrelated process. Many questions are still open on the composition, molecular interactions, assembly pathway, mechanism of action, and physiological significance of this molecular machine. To get further insights into the structure and function of this protein complex, we investigated on the properties of RNA degradosomes purified from pnp mutant strains expressing PNPases with single aminoacid substitutions in different domains. None of mutations tested impaired assembly of PNPase with RNase E and, for most mutants, the RNA-degrading ability of the degradosome correlated with the phosphorolytic activity of the mutant PNPase. An exception was Pnp-E81D, which harbors a single aminoacid substitution on the outer side of the trimerization interface. Although in crude extracts neither phosphorolytic activity nor RNA binding appeared to be impaired, the mutant was defective in autogenous regulation. The RNA degradosome containing Pnp-E81D was unable to degrade a structured RNA substrate. This seemed to correlate with a lower abundance of the RhlB RNA helicase. It thus appears that PNPase participates in the assembly of RhlB in the degradosome and/or in the control of the RNA helicase activity. In addition, these data suggest the participation of the RhlB RNA helicase in PNPase autogenous regulation.
|Titolo:||Characterization of RNA degradosome from E. coli with mutant polynucleotide phosphorylase|
DEHO', GIOVANNI (Ultimo)
|Data di pubblicazione:||2006|
|Settore Scientifico Disciplinare:||Settore BIO/18 - Genetica|
|Enti collegati al convegno:||Societa’ Italiana di Microbiologia Generale e Biotecnologie Microbiche|
|Citazione:||Characterization of RNA degradosome from E. coli with mutant polynucleotide phosphorylase / M. Del Favero, E. Mazzantini, F. Briani, P. Tortora, G. Dehò. ((Intervento presentato al 25. convegno Convegno Nazionale SIMGBM Societa’ Italiana di Microbiologia Generale e Biotecnologie Microbiche tenutosi a Orvieto nel 2006.|
|Appare nelle tipologie:||14 - Intervento a convegno non pubblicato|