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Proteome-Wide Alterations in Translation Rates of E. coli Under Anaerobic Conditions

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asher
10 diapositives · 272 Views · Aug 04, 2014

This study investigates the impact of anaerobiosis on translation rates and protein synthesis in Escherichia coli. Using combined fractional diagonal chromatography (COFRADIC) and a pulse-labeling technique with the methionine analogue azidohomoalanine (azhal), we observed a rapid increase in synthesis rates for proteins related to anaerobic respiration and fermentation after just 10 minutes of anaerobiosis. Additionally, the synthesis rates of glycolytic enzymes and PTS proteins were enhanced, highlighting the interplay between metabolic regulation and protein synthesis at the ribosomal level. Key findings suggest that proteomic changes occur independently of mRNA degradation.

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Proteome-Wide Alterations in Translation Rates of E. coli Under Anaerobic Conditions
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  1. mRNA Degradation vs Translation

  2. Proteome-wide alterations in Escherichia coli translation rates upon anaerobiosis 2010

  3. Combined fractional diagonal chromatography • COFRADIC enriched azidohomoalanine (azhal) containing peptides using tris-(2-carboxy-ethyl)-phosphine (TCEP) to selectively modify peptides • Developed a pulse labeling method using the methionine analogue azhal • Tested after 10 minutes of anaerobiosis

  4. Immediate and strong increase in synthesis rate of proteins involved in anaerobic respiration and fermentation. • The synthesis rates of glycolytic enzymes and PTS proteins increased as well, suggesting that the increased flux through glycolysis is not exclusively the result of metabolic regulation, but also regulated at the protein level. • Most of the proteins found to be down-regulated in synthesis were ribosomal proteins. • It has been well documented that under conditions of energy deficiency synthesis of rRNA is decreased leading to translational feedback inhibition of ribosomal proteins synthesis • These changes are induced by small regulatory RNAs that affect translation

  5. Changes in the proteome can occur independently of mRNA degradation

  6. Poly(A)-polymerase I links transcription with mRNA degradation via σSproteolysis 2006

  7. Figure 6. PAPI-mediated effect on σS proteolysis. A. • σSis expressed under stressful conditions (stationary phase). PAPI reduces σSproteolysis under stressful conditions as shown by the decreased σS in the mutant without PAPI (two columns on the right of each graph and the right side of the blot)

  8. Figure 8. Interplay among transcription, degradation and proteolysis machineries in the control of BolA and σSBolA levels in the cell • PAPI reduces proteolysis of σScaused by RssB and ClpX and reduces mRNA degradation caused by various enzymes. The increased σS levels induce the transcription and translation of various other proteins

  9. mRNA degradation, proteolysis, and transcription work in concert to control subsequent gene expression and protein generation

  10. Papers • http://www.ncbi.nlm.nih.gov/pubmed/20713451 • http://www.ncbi.nlm.nih.gov/pubmed/16556229