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J Bacteriol, April 1998, p. 1759-1765, Vol. 180, No. 7
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Genetic Complementation and Kinetic Analyses of
Rhodobacter capsulatus ORF1696 Mutants Indicate that the
ORF1696 Protein Enhances Assembly of the Light-Harvesting I
Complex
C. S.
Young,
R. C.
Reyes, and
J. T.
Beatty*
Department of Microbiology and Immunology,
The University of British Columbia, Vancouver, British Columbia,
Canada V6T 1Z3
Received 23 October 1997/Accepted 19 January 1998
Rhodobacter capsulatus ORF1696 mutant strains were
created by insertion of antibiotic resistance cartridges at different
sites within the ORF1696 gene in a strain that lacks the
light-harvesting II (LHII) complex. Steady-state absorption
spectroscopy profiles and the kinetics of the light-harvesting I (LHI)
complex assembly and decay were used to evaluate the function of the
ORF1696 protein in various strains. All of the mutant strains were
found to be deficient in the LHI complex, including one (
Nae) with a
disruption located 13 codons before the 3' end of the gene. A
5'-proximal disruption after the 31st codon of ORF1696
resulted in a mutant strain (
Mun) with a novel absorption spectrum.
The two strains with more 3' disruptions (
Stu and
Nae) were
restored nearly to the parental strain phenotype when trans
complemented with a plasmid expressing the ORF1696 gene,
but
Mun was not. The absorption spectrum of
Mun resembled that of
a strain which had a polar mutation in ORF1696. We suggest
that a rho-dependent transcription termination site exists
between the MunI and proximal StuI sites of
ORF1696. A comparison of LHI complex assembly kinetics
showed that assembly occurred 2.6-fold faster in the parental strain than in strain
Stu. In contrast, LHI complex decay occurred 1.7-fold faster in the ORF1696 parental strain than in
Stu. These
results indicate that the ORF1696 protein has a major effect on LHI
complex assembly, and models of ORF1696 function are proposed.
*
Corresponding author. Mailing address: Department of
Microbiology & Immunology, The University of British Columbia, Rm. 300, 6174 University Blvd., Vancouver, BC, Canada V6T 1Z3. Phone: (604) 822-6896. Fax: (604) 822-6041. E-mail:
jbeatty{at}unixg.ubc.ca.
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