The purpose of this proposal is to understand the relationship between membrane architecture and function in photosynthetic membranes. Our approach will be via discrete, yet correlative, techniques using the unicellular, transformable cyanobacterium Anacystis nidulans R2. The emphasis will be on Photosystem II and the oxygen-evolving apparatus. Our strategy will consist of seven major experimental techniques: (1) the isolation and characterization of an oxygen-evolving complex that contains Photosystem II and phycobilisomes; (2) site-specific labeling and cross-linking of proteins to determine their position in the membrane; (3) reconstitution of photosynthetic electron transport from H2O to NADP+ in liposomes using isolated complexes; (4) isolation of photosynthetic mutants and herbicide-resistant mutants. In addition to temperature-sensitive mutants, strains will be isolated that must be maintained by heterotrophic growth; (5) complementation analysis with cloned genes; (6) cloned photosynthesis genes will be used for biochemical and genetic experiments; and (7) fluorescence analysis of chlorophyllprotein complexes in normal and developing cells. This project is a combined genetic and biochemical study of membrane structure and function and is thus of great benefit to membrane biology in general. The specific information that should be obtained from this proposal includes: (1) the protein composition of the major membrane complexes, such as the O2-evolving apparatus and chlorophyll-proteins; (2) the precise location of proteins in the membrane; (3) the effects of mutation on membrane structure and function; (4) the effect of adding cloned photosynthesis genes to wild-type and mutant cells; (5) the identity of genes coding for photosynthetic proteins; (6) the identity of the chlorophyll components that give rise to the 77 K fluorescence emission peaks; and (7) the mechanism of membrane assembly in iron-deficient cells after iron addition and in heterotrophic cells after light induction. The combination of biochemistry and genetics makes A. nidulans a very valuable organism for the study of bioenergetics and membrane structure.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM021827-11
Application #
3270724
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1978-05-01
Project End
1989-04-30
Budget Start
1985-05-01
Budget End
1986-04-30
Support Year
11
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of Missouri-Columbia
Department
Type
Schools of Arts and Sciences
DUNS #
112205955
City
Columbia
State
MO
Country
United States
Zip Code
65211
Bullerjahn, G S; Jensen, T C; Sherman, D M et al. (1990) Immunological characterization of the Prochlorothrix hollandica and Prochloron sp. chlorophyll a/b antenna proteins. FEMS Microbiol Lett 55:99-105
Webb, R; Reddy, K J; Sherman, L A (1990) Regulation and sequence of the Synechococcus sp. strain PCC 7942 groESL operon, encoding a cyanobacterial chaperonin. J Bacteriol 172:5079-88
Reddy, K J; Masamoto, K; Sherman, D M et al. (1989) DNA sequence and regulation of the gene (cbpA) encoding the 42-kilodalton cytoplasmic membrane carotenoprotein of the cyanobacterium Synechococcus sp. strain PCC 7942. J Bacteriol 171:3486-93
Webb, R; Reddy, K J; Sherman, L A (1989) Lambda ZAP: improved strategies for expression library construction and use. DNA 8:69-73
Riethman, H C; Mawhinney, T P; Sherman, L A (1988) Characterization of phycobilisome glycoproteins in the cyanobacterium Anacystis nidulans R2. J Bacteriol 170:2433-40
Reddy, K J; Bullerjahn, G S; Sherman, D M et al. (1988) Cloning, nucleotide sequence, and mutagenesis of a gene (irpA) involved in iron-deficient growth of the cyanobacterium Synechococcus sp. strain PCC7942. J Bacteriol 170:4466-76
Riethman, H C; Sherman, L A (1988) Purification and characterization of an iron stress-induced chlorophyll-protein from the cyanobacterium Anacystis nidulans R2. Biochim Biophys Acta 935:141-51
Kuwabara, T; Reddy, K J; Sherman, L A (1987) Nucleotide sequence of the gene from the cyanobacterium Anacystis nidulans R2 encoding the Mn-stabilizing protein involved in photosystem II water oxidation. Proc Natl Acad Sci U S A 84:8230-4
Bullerjahn, G S; Matthijs, H C; Mur, L R et al. (1987) Chlorophyll-protein composition of the thylakoid membrane from Prochlorothrix hollandica, a prokaryote containing chlorophyll b. Eur J Biochem 168:295-300
Riethman, H C; Mawhinney, T P; Sherman, L A (1987) Phycobilisome-associated glycoproteins in the cyanobacterium Anacystis nidulans R 2. FEBS Lett 215:209-14

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