These investigations will focus on using molecular biological, biochemical, and genetic methods to elucidate how messenger RNA is degraded in bacteria. Particular attention will be devoted to RppH, a recently discovered RNA pyrophosphohydrolase that triggers the degradation of primary transcripts in E. coli by a previously unrecognized mechanism: the rate-determining conversion of the 5'-terminal triphosphate to a monophosphate. First, the features of RNAs that determine their susceptibility to decay by this mechanism will be identified. In addition, the characteristics of RppH that are important for its activity and the cellular factors that may influence such 5'-end- dependent RNA degradation will be examined. Finally, the contribution of this pathway to mRNA decay in other bacterial species will be explored. The results of these studies will provide important insights into a fundamental aspect of gene regulation that presently is poorly understood. This knowledge should be of value in clarifying a biological regulatory mechanism that can play a key role in bacterial pathogenesis.

Public Health Relevance

The proposed research will address the mechanism of bacterial messenger RNA degradation, a basic biological process important for controlling gene expression in all living organisms. The knowledge thereby acquired should be of value in maximizing bacterial production of medically useful proteins and in elucidating a regulatory mechanism that can influence bacterial pathogenesis.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM035769-26S1
Application #
8669324
Study Section
Program Officer
Bender, Michael T
Project Start
2013-09-01
Project End
2015-08-31
Budget Start
2013-09-01
Budget End
2015-08-31
Support Year
26
Fiscal Year
2013
Total Cost
$80,004
Indirect Cost
$32,804
Name
New York University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
121911077
City
New York
State
NY
Country
United States
Zip Code
10016
Vogel, Jörg; Gottesman, Susan; Belasco, Joel et al. (2014) Meeting report: Regulating with RNA in Bacteria 2013. RNA Biol 11:403-12
Hsieh, Ping-kun; Richards, Jamie; Liu, Quansheng et al. (2013) Specificity of RppH-dependent RNA degradation in Bacillus subtilis. Proc Natl Acad Sci U S A 110:8864-9
Richards, Jamie; Liu, Quansheng; Pellegrini, Olivier et al. (2011) An RNA pyrophosphohydrolase triggers 5'-exonucleolytic degradation of mRNA in Bacillus subtilis. Mol Cell 43:940-9
Richards, Jamie; Belasco, Joel G (2011) Ribonuclease J: how to lead a double life. Structure 19:1201-3
Yao, Shiyi; Richards, Jamie; Belasco, Joel G et al. (2011) Decay of a model mRNA in Bacillus subtilis by a combination of RNase J1 5' exonuclease and RNase Y endonuclease activities. J Bacteriol 193:6384-6
Belasco, Joel G (2010) All things must pass: contrasts and commonalities in eukaryotic and bacterial mRNA decay. Nat Rev Mol Cell Biol 11:467-78
Schuck, Alyssa; Diwa, Alexis; Belasco, Joel G (2009) RNase E autoregulates its synthesis in Escherichia coli by binding directly to a stem-loop in the rne 5'untranslated region. Mol Microbiol 72:470-8
Messing, Simon A J; Gabelli, Sandra B; Liu, Quansheng et al. (2009) Structure and biological function of the RNA pyrophosphohydrolase BdRppH from Bdellovibrio bacteriovorus. Structure 17:472-81
Celesnik, Helena; Deana, Atilio; Belasco, Joel G (2008) PABLO analysis of RNA 5'-phosphorylation state and 5'-end mapping. Methods Enzymol 447:83-98
Celesnik, Helena; Deana, Atilio; Belasco, Joel G (2007) Initiation of RNA decay in Escherichia coli by 5'pyrophosphate removal. Mol Cell 27:79-90

Showing the most recent 10 out of 33 publications