The relationship between the trimethylation of lysine 77 and cytochrome c metabolism in Saccharomyces cerevisiae.
1. Site directed mutations were constructed in the yeast iso-1-cytochrome c gene adjacent to the lysine 77 (methylation site) codon. 2. These mutant genes were then cloned and transformed into the S. cerevisiae strain B-6642 which contains a deficiency in the iso-1-cytochrome c gene. 3. The resulting transformants were screened for cytochrome c production using gel electrophoresis. 4. Amino acid analysis of the mutated cytochromes c demonstrated varying levels of trimethyllysine formation, depending on the nature of the site directed mutation. 5. The resulting transformants were then used as tools in order to investigate the relationship between trimethyllysine formation and various aspects of cytochrome c metabolism including protein stability and heme conjugation.
Ceesay
K J
KJ
Department of Chemistry, Ohio University, Athens 45701.
Rider
L R
LR
Bergman
L W
LW
Tuck
M T
MT
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D003574:Cytochrome c Group / Q000235:genetics / Q000378:metabolism*; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004355:Drug Stability; D018014:Gene Transfer Techniques; D006418:Heme / Q000378:metabolism; D008239:Lysine / Q000031:analogs & derivatives* / Q000378:metabolism*; D008745:Methylation; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*
D003574:Cytochrome c Group; C003712:trimethyllysine; D006418:Heme; D004262:DNA Restriction Enzymes; D008239:Lysine