Novel gene expression mechanism in a fission yeast retroelement: Tf1 proteins are derived from a single primary translation product.
In sharp contrast to the single ORF of the Schizosaccharomyces pombe retrotransposon Tf1, retroviruses and most retrotransposons employ two different ORFs to separately encode the Gag and Pol proteins. The different ORFs are thought to allow for overexpression of the Gag protein relative to Pol protein presumed necessary for the assembly of functional retrovirus particles and virus-like particles (VLPs). The results of in vivo experiments designed to detect the transposition of Tf1 show that Tf1 is indeed active and can insert itself into the host genome via a true retrotransposition process. Thus, a paradox emerged between the lack of any obvious means of overexpressing Tf1 Gag protein and the demonstrated functionality of the element. Epitope tagging experiments described here confirm that the Tf1 large ORF is intact and that there is no translational or transcriptional mechanism used to overexpress the Tf1 Gag protein. In addition, we used sucrose gradients and antisera specific for Tf1 capsid (CA) and integrase (IN) to show that the Tf1 proteins do assemble into uniform populations of macromolecular particles that also cosediment with Tf1 reverse transcription products. This evidence suggests that Tf1 proteins form VLPs without using the previously described mechanisms that retroviruses and retrotransposons require to overexpress Gag proteins.
Levin
H L
HL
Department of Molecular Biology and Genetics, Johns Hopkins School of Medicine, Baltimore, MD 21205.
Weaver
D C
DC
Boeke
J D
JD
D001483:Base Sequence; D004251:DNA Transposable Elements; D004271:DNA, Fungal; D005656:Fungal Proteins / Q000096:biosynthesis / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D015683:Gene Products, gag / Q000096:biosynthesis / Q000235:genetics; D015151:Immunoblotting; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D012568:Schizosaccharomyces / Q000235:genetics*; D013395:Sucrose; D014158:Transcription, Genetic
D004251:DNA Transposable Elements; D004271:DNA, Fungal; D005656:Fungal Proteins; D015683:Gene Products, gag; D013395:Sucrose