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Cloning and characterization of a cDNA encoding Xenopus laevis alpha o1 isoform of the Go protein
L Paquereau1, E Devic, Y Audigier
1UMR 9925, Universite Toulouse-III, France.
Abstract:
The mammalian gene encoding the alpha subunit of the Go protein generates by alternative splicing two isoforms, alpha o1 and alpha o2, which differ in their carboxy-terminal region. We report here the cloning of a Xenopus cDNA (XG alpha o1) which encodes a protein corresponding to the mammalian alpha o1 isoform. In its 3' untranslated region, the transcript contains a repetitive motif made up of dinucleotide AT repeats. By RT-PCR amplification, we showed that XG alpha o1 transcripts are both maternal and zygotic. As alpha o2 transcripts have been shown to be maternal and devoid of AT repeats, the repetitive motif could play a role in the differential expression of each isoform.
Insights
Researchers cloned a Xenopus gene (XG alpha o1) encoding a Go protein alpha subunit isoform. Its transcript contains AT repeats, potentially regulating differential expression compared to other isoforms.
Area of Science:
- Molecular Biology
- Developmental Biology
- Xenopus laevis Research
Background:
- The Go protein alpha subunit gene produces two isoforms, alpha o1 and alpha o2, via alternative splicing.
- These isoforms differ in their carboxy-terminal regions, suggesting distinct functional roles.
Purpose of the Study:
- To clone and characterize the Xenopus homolog of the mammalian alpha o1 Go protein subunit.
- To investigate the regulatory elements within the XG alpha o1 transcript.
Main Methods:
- Cloning of Xenopus cDNA (XG alpha o1).
- Analysis of the 3' untranslated region for repetitive motifs.
- RT-PCR amplification to determine transcript expression patterns.
Main Results:
- A Xenopus cDNA (XG alpha o1) encoding a protein similar to the mammalian alpha o1 isoform was successfully cloned.
- The XG alpha o1 transcript's 3' untranslated region contains a repetitive motif of AT dinucleotides.
- RT-PCR confirmed that XG alpha o1 transcripts are present in both maternal and zygotic stages.
Conclusions:
- The cloned XG alpha o1 represents a functional homolog of the mammalian alpha o1 isoform.
- The identified AT-rich motif in the 3' UTR may be involved in the differential expression of XG alpha o1.
- Comparing XG alpha o1 with alpha o2 transcripts suggests a regulatory mechanism for isoform-specific gene expression in Xenopus development.