Messenger RNA turnover has long been recognized to play a key role in controlling gene expression in all organisms. The long-term goal of this research project is to understand the basic principles that govern mRNA degradation in bacterial cells. The immediate objectives are to elucidate the specificity and mechanism of 5'-end-dependent RNA degradation in Escherichia coli and Bacillus subtilis by investigating the proteins that are central to this regulatory pathway and the transcripts that they target. Achieving these objectives will require not only the use of standard molecular biological, biochemical, and genetic methods but also the development of new low- and high-throughput techniques for analyzing the 5' phosphorylation state of individual transcripts and for using genetic screening to identify pertinent clones and mutants. The knowledge gained from these studies will provide important insights into a fundamental aspect of gene regulation that can play a key role in bacterial pathogenesis.

Public Health Relevance

The proposed research will address the mechanisms by which messenger RNA is degraded in bacterial cells. The knowledge thereby acquired is expected to be of value in understanding the regulatory processes that govern bacterial pathogenesis and in maximizing bacterial production of medically useful proteins. In addition, the methods and concepts developed in the course of these studies are likely to be useful for elucidating how messenger RNA degradation helps to ensure proper levels of gene expression in healthy human cells.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Prokaryotic Cell and Molecular Biology Study Section (PCMB)
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Bender, Michael T
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New York University
Schools of Medicine
New York
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Luciano, Daniel J; Vasilyev, Nikita; Richards, Jamie et al. (2018) Importance of a diphosphorylated intermediate for RppH-dependent RNA degradation. RNA Biol 15:703-706
Luciano, Daniel J; Belasco, Joel G (2018) Analysis of RNA 5' ends: Phosphate enumeration and cap characterization. Methods :
Gao, Ang; Vasilyev, Nikita; Luciano, Daniel J et al. (2018) Structural and kinetic insights into stimulation of RppH-dependent RNA degradation by the metabolic enzyme DapF. Nucleic Acids Res 46:6841-6856
Belasco, Joel G (2017) Death by translation: ribosome-assisted degradation of mRNA by endonuclease toxins. FEBS Lett 591:1851-1852
Bischler, Thorsten; Hsieh, Ping-Kun; Resch, Marcus et al. (2017) Identification of the RNA Pyrophosphohydrolase RppH of Helicobacter pylori and Global Analysis of Its RNA Targets. J Biol Chem 292:1934-1950
Lodato, Patricia B; Thuraisamy, Thujitha; Richards, Jamie et al. (2017) Effect of RNase E deficiency on translocon protein synthesis in an RNase E-inducible strain of enterohemorrhagic Escherichia coli O157:H7. FEMS Microbiol Lett 364:
Luciano, Daniel J; Vasilyev, Nikita; Richards, Jamie et al. (2017) A Novel RNA Phosphorylation State Enables 5' End-Dependent Degradation in Escherichia coli. Mol Cell 67:44-54.e6
Richards, Jamie; Belasco, Joel G (2016) Distinct Requirements for 5'-Monophosphate-assisted RNA Cleavage by Escherichia coli RNase E and RNase G. J Biol Chem 291:5038-48
Belasco, Joel G (2015) Way to go, RNA. RNA 21:565-6
Luciano, Daniel J; Belasco, Joel G (2015) NAD in RNA: unconventional headgear. Trends Biochem Sci 40:245-7

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