Abstract
ORF3 of the cytoplasmic linear plasmid pGKL2 was disrupted in vivo by integration of a selectable marker. Long-term cultivation of transformants carrying hybrid plasmids with a disrupted ORF3 under selective pressure did not deprive strains of the native counterpart, thereby proving its essentiality for pGKL2 replication and maintenance. The predicted ORF3 polypeptide was found to contain conserved motifs acquainted with mRNA-capping enzymes in the required order, just as in cytoplasmic viruses; new conserved motifs were also identified.
Copyright 1998 Academic Press.
MeSH terms
- Acid Anhydride Hydrolases / genetics*
- Acid Anhydride Hydrolases / isolation & purification
- Amino Acid Sequence
- DNA, Fungal / genetics*
- Fungal Proteins / genetics*
- Fungal Proteins / isolation & purification
- Genes, Fungal*
- Kluyveromyces / genetics*
- Methyltransferases / genetics*
- Methyltransferases / isolation & purification
- Molecular Sequence Data
- Nucleotidyltransferases / genetics*
- Nucleotidyltransferases / isolation & purification
- Open Reading Frames
- Plasmids / genetics
- Plasmids / physiology*
- RNA Caps / metabolism
- RNA Processing, Post-Transcriptional
- RNA, Fungal / metabolism
- RNA, Messenger / metabolism
- Sequence Alignment
- Sequence Homology, Amino Acid
Substances
- DNA, Fungal
- Fungal Proteins
- RNA Caps
- RNA, Fungal
- RNA, Messenger
- Methyltransferases
- Nucleotidyltransferases
- mRNA capping enzyme, Kluyveromyces lactis
- Acid Anhydride Hydrolases