The targeting and assembly of proteins are fundamental biological processes. In many instances, the targeting signal is defined by small liner stretches of sequence such as those present at the amino terminal end of secreted proteins in bacteria and eukaryotes. While these sequences in Escherichia coli assist proteins in exiting the cytoplasm, they do not determine the final location of secreted proteins within the extracytoplasmic compartments: periplasm and outer membrane. Results from this project have emphasized the view that the targeting signal for outer membrane proteins resides within the partially folded intermediates. The folding states of these intermediates are guided by the intrinsic properties of the molecule and its interactions with other cellular components including chaperone proteins and lipopolysaccharide, which is exclusively localized in the outer membrane. Novel genetic approaches outlined in this project will assist in revealing the nature of intragenic information and provide a better understanding of the dynamic interactions between various cellular components during the targeting and assembly of outer membrane proteins. Because interactions between lipidic components and proteins occur in all biological membranes, the results should have broad general interest. OmpF, OmpC and LamB will be used as model proteins. These proteins serve as the receptor for bacteriophages and form channels that allow the diffusion of water soluble solutes. Three-dimensional structures of OmpF and LamB have been resolved which, while permitting a better understanding of the structure-function relationship of these proteins, shed little light on how they reach their destination. The long term goals of this project are to understand the biogenesis of outer membrane proteins and molecular architecture of the outer membrane. These objectives will be achieved through the application of genetic approaches that are feasible and have already yielded important and new insights.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM048167-07
Application #
2749925
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1992-08-01
Project End
2001-07-31
Budget Start
1998-08-01
Budget End
1999-07-31
Support Year
7
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Arizona State University-Tempe Campus
Department
Microbiology/Immun/Virology
Type
Schools of Arts and Sciences
DUNS #
188435911
City
Tempe
State
AZ
Country
United States
Zip Code
85287
Misra, Rajeev; Stikeleather, Ryan; Gabriele, Rebecca (2015) In vivo roles of BamA, BamB and BamD in the biogenesis of BamA, a core protein of the ?-barrel assembly machine of Escherichia coli. J Mol Biol 427:1061-74
Workman, Patricia; Heide, Kristina; Giuliano, Nicolas et al. (2012) Genetic, biochemical, and molecular characterization of the polypeptide transport-associated domain of Escherichia coli BamA. J Bacteriol 194:3512-21
Leiser, Owen P; Charlson, Emily S; Gerken, Henri et al. (2012) Reversal of the ?degP phenotypes by a novel rpoE allele of Escherichia coli. PLoS One 7:e33979
Leonard-Rivera, Margaret; Misra, Rajeev (2012) Conserved residues of the putative L6 loop of Escherichia coli BamA play a critical role in the assembly of ýý-barrel outer membrane proteins, including that of BamA itself. J Bacteriol 194:4662-8
Tellez Jr, Rene; Misra, Rajeev (2012) Substitutions in the BamA ?-barrel domain overcome the conditional lethal phenotype of a ?bamB ?bamE strain of Escherichia coli. J Bacteriol 194:317-24
Bennion, Drew; Charlson, Emily S; Coon, Eric et al. (2010) Dissection of ýý-barrel outer membrane protein assembly pathways through characterizing BamA POTRA 1 mutants of Escherichia coli. Mol Microbiol 77:1153-71
Gerken, Henri; Misra, Rajeev (2010) MzrA-EnvZ interactions in the periplasm influence the EnvZ/OmpR two-component regulon. J Bacteriol 192:6271-8
Gerken, Henri; Leiser, Owen P; Bennion, Drew et al. (2010) Involvement and Necessity of the Cpx Regulon in the Event of Aberrant beta-barrel Outer Membrane Protein Assembly. Mol Microbiol :
Gerken, Henri; Charlson, Emily S; Cicirelli, Elisha M et al. (2009) MzrA: a novel modulator of the EnvZ/OmpR two-component regulon. Mol Microbiol 72:1408-22
Masi, Muriel; Duret, Guillaume; Delcour, Anne H et al. (2009) Folding and trimerization of signal sequence-less mature TolC in the cytoplasm of Escherichia coli. Microbiology 155:1847-57

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