A spontaneous tRNA suppressor of a mutation in the Chlamydomonas reinhardtii nuclear MCD1 gene required for stability of the chloroplast petD mRNA.
Numerous nuclear gene products are required for the correct expression of organellar genes. One such gene in the green alga Chlamydomonas reinhardtii is MCD1, whose product is required for stability of the chloroplast-encoded petD mRNA. In mcd1 mutants, which are non-photosynthetic, petD mRNA is degraded by a 5'-3' exonuclease activity, resulting in a failure to synthesize its product, subunit IV of the cytochrome b (6)/f complex. Here, we report the sequence of the wild-type MCD1 gene, which encodes a large and novel putative protein. Analysis of three mutant alleles showed that two harbored large deletions, but that one allele, mcd1-2, had a single base change resulting in a nonsense codon near the N-terminus. This same mutant allele can be suppressed by a second-site mutation in the nuclear MCD2 gene, whereas mcd2-1 cannot suppress the deletion in mcd1-1 (Esposito,D. Higgs,D.C. Drager,R.G. Stern, D.B. and Girard-Bascou,J. (2001) Curr. Genet., 39, 40-48). We report the cloning of mcd2-1, and show that the mutation lies in a tRNA(Ser)(CGA), which has been modified to translate the nonsense codon in mcd1-2. We discuss how the existence of a large tRNA(Ser) gene family may permit this suppression without pleiotropic consequences.
Murakami
Shinya
S
Boyce Thompson Institute for Plant Research, Cornell University Tower Road, Ithaca, NY 14853, USA.
Kuehnle
Katrin
K
Stern
David B
DB
D020418:Algal Proteins / Q000235:genetics; D000483:Alleles; D000818:Animals; D001483:Base Sequence; D002467:Cell Nucleus / Q000235:genetics; D016825:Chlamydomonas reinhardtii / Q000235:genetics* / Q000378:metabolism; D002736:Chloroplasts / Q000235:genetics; D003001:Cloning, Molecular; D045346:Cytochrome b6f Complex / Q000235:genetics* / Q000378:metabolism; D016153:Genes, Suppressor; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D020871:RNA Stability; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism*; D012362:RNA, Transfer, Ser / Q000235:genetics* / Q000378:metabolism; D013489:Suppression, Genetic
D020418:Algal Proteins; D012333:RNA, Messenger; D012362:RNA, Transfer, Ser; C084072:petD protein, cytochrome b(6)f complex; D045346:Cytochrome b6f Complex