Isolation and characterization of a Chlamydomonas reinhardtii mutant lacking the gamma-subunit of chloroplast coupling factor 1 (CF1).
Several nonphotoautotrophic mutants of Chlamydomonas reinhardtii were generated by transforming strain nit1-305 (cw 15) with exogenous DNA. An enrichment for potential photophosphorylation mutants was performed on medium containing arsenate, and acetate-requiring auxotrophs were then identified by replica plating. Strains containing a potential mutation in the nuclear DNA encoding the chloroplast coupling factor 1 (CF1) gamma-subunit (the atpC gene) were first identified serologically with a monospecific antiserum directed against the CF1 gamma-subunit polypeptide. Of several mutants isolated, one, designated T1-54, was characterized at the protein, DNA, and RNA levels. Mutant strain T1-54 lacks anti-CF1 gamma-subunit cross-reacting material, exhibits polymorphism at the atpC locus compared with the parental strain, and lacks the mRNA transcript for the CF1 gamma-subunit. The data are consistent with there being an insertion of exogenous DNA, a deletion of DNA, or both at the 5' end of the gene encoding the CF1 gamma-subunit.
Smart
E J
EJ
Department of Biochemistry, College of Agricultural and Life Sciences, University of Wisconsin-Madison 53706.
Selman
B R
BR
D000085:Acetates / Q000378:metabolism; D000818:Animals; D001149:Arsenates / Q000378:metabolism; D015152:Blotting, Northern; D015139:Blotting, Southern; D002696:Chlamydomonas / Q000235:genetics* / Q000302:isolation & purification; D002736:Chloroplasts / Q000378:metabolism; D002872:Chromosome Deletion; D015151:Immunoblotting; D009154:Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D006180:Proton-Translocating ATPases / Q000235:genetics*; D014170:Transformation, Genetic
D000085:Acetates; D001149:Arsenates; D006180:Proton-Translocating ATPases; C025657:arsenic acid