Formation of C-terminally truncated version of the Taz1 protein employs cleavage-box structure in mRNA.
When expressed in various hosts the taz1(+) gene encoding the fission yeast telomere-binding protein produces two forms of polypeptides: full-length (Taz1p) and truncated (Taz1pDeltaC) version lacking almost entire Myb-domain. Whereas Taz1p binds telomeric DNA in vitro, Taz1pDeltaC forms long filaments unable of DNA binding. The formation of Taz1pDeltaC is a result of neither site-specific proteolysis, nor premature termination of transcription. In silico analysis of the taz1(+) RNA transcript revealed a stem-loop structure at the site of cleavage (cleavage box; CB). In order to explore whether it possesses inherent destabilizing effects, we cloned CB sequence into the open reading frame (ORF) of glutathione-S-transferase (GST) and observed that when expressed in Escherichia coli the engineered gene produced two forms of the reporter protein. The formation of the truncated version of GST was abolished, when CB was replaced with recoded sequence containing synonymous codons thus indicating that the truncation is based on structural properties of taz1(+) mRNA.
Gunisova
Stanislava
S
Department of Genetics, Comenius University, 842 15 Bratislava, Slovakia.
Bartosova
Zdenka
Z
Kramara
Juraj
J
Nosek
Jozef
J
Tomaska
Lubomir
L
D000071183:Autophagy-Related Proteins; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism*; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D009690:Nucleic Acid Conformation; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism*; D017423:Sequence Analysis, RNA; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic
D000071183:Autophagy-Related Proteins; D002868:Chromosomal Proteins, Non-Histone; D012333:RNA, Messenger; D029702:Schizosaccharomyces pombe Proteins; D014157:Transcription Factors; C513475:atg11 protein, S pombe; D005982:Glutathione Transferase