The machinery responsible for making proteins (e.g. ribosomal RNA, ribosomal proteins, translation factors, and tRNAs) is central to growth and development of all organisms. The control of its synthesis has been a central issue in molecular microbiolgy for almost 60 years. It had become clear in previous project periods that our work was central not only for understanding ribosome synthesis but also for understanding the mechanism of transcription initiation in general. What has become apparent in the last few years is that the systems that regulate rRNA promoters also directly regulate a wide variety of other promoters as well, many of which were not anticipated because their connection to the protein synthetic apparatus was not apparent. However, we now know that the molecules that regulate rRNA promoters are utilized throughout the bacterial kingdom, they play crucial roles in virulence and infectious disease, and they contribute to genome maintenance as well as to transcription. In the next project period, we will (i) continue our studies on the DNA-RNAP interactions that contribute to the transition from a closed to an open complex in the promoter mechanism, (ii) explore the role of specific promoter regions in defining the transcription start site, (iii) continue our examination of the control of transcription by ppGpp and DksA by defining their binding sites on RNA polymerase and determining their mechanism of action, (iv) expand our analysis of control by these molecules to additional promoters in order to understand what differentiates positively from negatively regulated targets, (v) continue studying the mechanism of action ofthe RNA polymerase assembly factor CrI, and (vi) continue studies on our recent discovery that E. coli makes a structure analogous to the eukaryotic nucleolus. We will expand these studies to explore other potential long-range interactions between distant parts of the bacterial chromosome.

Public Health Relevance

It has become apparent in the lastfew years that the systems that regulate rRNA promoters also directly regulate a wide variety of other promoters as well. Some of these factors play crucial roles in virulence and infectious disease, as well as contributing to gene expression and the fidelity of replication. Thus, understanding these fundamental processes has the potential to impact human health directly.

Agency
National Institute of Health (NIH)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
3R37GM037048-28S1
Application #
8830183
Study Section
No Study Section (in-house review) (NSS)
Program Officer
Sledjeski, Darren D
Project Start
Project End
Budget Start
Budget End
Support Year
28
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Microbiology/Immun/Virology
Type
Earth Sciences/Resources
DUNS #
City
Madison
State
WI
Country
United States
Zip Code
53715
Lennon, Christopher W; Lemmer, Kimberly C; Irons, Jessica L et al. (2014) A Rhodobacter sphaeroides protein mechanistically similar to Escherichia coli DksA regulates photosynthetic growth. MBio 5:e01105-14
Banta, Amy B; Cuff, Marianne E; Lin, Hueylie et al. (2014) Structure of the RNA polymerase assembly factor Crl and identification of its interaction surface with sigma S. J Bacteriol 196:3279-88
Banta, Amy B; Chumanov, Robert S; Yuan, Andy H et al. (2013) Key features of ýýS required for specific recognition by Crl, a transcription factor promoting assembly of RNA polymerase holoenzyme. Proc Natl Acad Sci U S A 110:15955-60
Winkelman, Jared T; Bree, Anna C; Bate, Ashley R et al. (2013) RemA is a DNA-binding protein that activates biofilm matrix gene expression in Bacillus subtilis. Mol Microbiol 88:984-97
My, Laetitia; Rekoske, Brian; Lemke, Justin J et al. (2013) Transcription of the Escherichia coli fatty acid synthesis operon fabHDG is directly activated by FadR and inhibited by ppGpp. J Bacteriol 195:3784-95
Ross, Wilma; Vrentas, Catherine E; Sanchez-Vazquez, Patricia et al. (2013) The magic spot: a ppGpp binding site on E. coli RNA polymerase responsible for regulation of transcription initiation. Mol Cell 50:420-9
Lee, Jeong-Hyun; Lennon, Christopher W; Ross, Wilma et al. (2012) Role of the coiled-coil tip of Escherichia coli DksA in promoter control. J Mol Biol 416:503-17
Chandrangsu, Pete; Wang, Li; Choi, Sang Ho et al. (2012) Suppression of a dnaKJ deletion by multicopy dksA results from non-feedback-regulated transcripts that originate upstream of the major dksA promoter. J Bacteriol 194:1437-46
Lemke, Justin J; Sanchez-Vazquez, Patricia; Burgos, Hector L et al. (2011) Direct regulation of Escherichia coli ribosomal protein promoters by the transcription factors ppGpp and DksA. Proc Natl Acad Sci U S A 108:5712-7
Chandrangsu, Pete; Lemke, Justin J; Gourse, Richard L (2011) The dksA promoter is negatively feedback regulated by DksA and ppGpp. Mol Microbiol 80:1337-48

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