The objective of this study is to characterize host-viral protein interactions involved in the expression of viral genes using coliphage Lambda and its E. coli host as a model system. The major emphasis will be directed towards analyzing the multiple factors involved in the regulation of gene expression by the N. transcription antitermination protein of Lambda. In addition to N, the components include the bacterial Nus proteins and a region on the phage genome, nut. We propose to continue our study of mutations that alter the activity of each of the components. Other possible components will be studied by characterizing mutations that suppress the effects of nus mutations. In particular, we will correlate structural differences with functional activity of the nusA gene and its product by analyzing: 1) conditionally lethal nusA mutations, 2) nusA genes from organisms other than E. coli, 3) hybrid E. coli-S. typhimurium nusA genes constructed in vitro, 4) truncated nusA proteins constructed by introducing chain terminating condons. The nut site will be studied to determine how sequence changes can create more optimal interactions with the component proteins. Mutations eliminating transcription termination will be analyzed to identify important elements in termination signals. A second set of E. coli mutations, sip, that influence the action of the lambdoid phage P22 cl transcription regulatory protein will be analyzed. The bacterial genes will be cloned and the proteins identified. Phage mutations that allow a bypass of the host block will be analyzed in order to determine the basis of the Sip defect. Studies on this well characterized system have significantly contributed to studies on the regulation of E. coli ribosomal RNA and biosynthetic operons. In the future they should also contribute to the study of other host-viral interactions such as those involved in malignant transformation.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI011459-16
Application #
3124947
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1976-09-01
Project End
1991-08-31
Budget Start
1988-09-01
Budget End
1989-08-31
Support Year
16
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Type
Schools of Medicine
DUNS #
791277940
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Tyler, Jessica S; Beeri, Karen; Reynolds, Jared L et al. (2013) Prophage induction is enhanced and required for renal disease and lethality in an EHEC mouse model. PLoS Pathog 9:e1003236
Bubunenko, Mikhail; Court, Donald L; Al Refaii, Abdalla et al. (2013) Nus transcription elongation factors and RNase III modulate small ribosome subunit biogenesis in Escherichia coli. Mol Microbiol 87:382-93
Friedman, David I; Mozola, Cara C; Beeri, Karen et al. (2011) Activation of a prophage-encoded tyrosine kinase by a heterologous infecting phage results in a self-inflicted abortive infection. Mol Microbiol 82:567-77
Eaton, Kathryn A; Friedman, David I; Francis, Gayle J et al. (2008) Pathogenesis of renal disease due to enterohemorrhagic Escherichia coli in germ-free mice. Infect Immun 76:3054-63
Tyler, Jessica S; Mills, Melissa J; Friedman, David I (2004) The operator and early promoter region of the Shiga toxin type 2-encoding bacteriophage 933W and control of toxin expression. J Bacteriol 186:7670-9
Livny, Jonathan; Friedman, David I (2004) Characterizing spontaneous induction of Stx encoding phages using a selectable reporter system. Mol Microbiol 51:1691-704
Tyler, Jessica S; Friedman, David I (2004) Characterization of a eukaryotic-like tyrosine protein kinase expressed by the Shiga toxin-encoding bacteriophage 933W. J Bacteriol 186:3472-9
Zhou, Y; Mah, T F; Yu, Y T et al. (2001) Interactions of an Arg-rich region of transcription elongation protein NusA with NUT RNA: implications for the order of assembly of the lambda N antitermination complex in vivo. J Mol Biol 310:33-49
Neely, M N; Friedman, D I (2000) N-mediated transcription antitermination in lambdoid phage H-19B is characterized by alternative NUT RNA structures and a reduced requirement for host factors. Mol Microbiol 38:1074-85
Huang, C; Wolfgang, M C; Withey, J et al. (2000) Charged tmRNA but not tmRNA-mediated proteolysis is essential for Neisseria gonorrhoeae viability. EMBO J 19:1098-107

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