Question

Alternative splicing within the chicken c-ets-1 locus: implications for transduction within the e26 retrovirus of the c-ets proto-oncogene.

Answer 1

A viral oncogene is created when an intensely transforming retrovirus, which causes tumors within weeks of infection, ingests genetic material from a host cell.

"Alternative splicing within the chicken c-ets-1 locus: implications for transduction within the e26 retrovirus of the c-ets proto-oncogene." Explain the experiment.

The alpha and beta genomic sequences corresponding to the 5' end of v-ets that were not found in the previously characterized c-ets RNA species or proteins were found in two overlapping c-ets-1 cDNA clones. These cDNAs were discovered to splice alpha and beta to a common set of 3' exons (a through F) already present in the p54c-ets-1 mRNA by nucleotide sequencing. They included a 1,455 nucleotide open reading frame that could decode a 485 amino acid polypeptide with a calculated molecular mass of 53 kilodaltons.

However, when expressed in COS-1 cells, the cDNAs directed the synthesis of a protein with an apparent molecular mass in sodium dodecyl sulfate-polyacrylamide gel electrophoresis of 68 kilodaltons, p68c-ets-1, comigrating with a protein expressed at low levels in normal chicken spleen cells. These two proteins were shown to be identical by partial digestion with protease V8. Northern (RNA) blot hybridization analysis with the p68c-its-1 -specific sequence and RNase protection experiments showed that the corresponding mRNA was expressed in normal chicken spleen and not in the normal chicken thymus or various T lymphoid cell lines.

As a result, two highly related proteins with differing amino-terminal regions are produced inside the same locus through the alternate attachment of several 5' exons, such as alpha and beta or I54, onto a common set of 3' exons (a to F). Finally, we show that the formation of the E26 myb-ets sequence involved an abnormal splicing event between a cryptic splice donor site in c-myb exon E6 and the usual splice acceptor site of c-ets-1 exon alpha.

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