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Related Experiment Videos

Alternatively spliced murine lyn mRNAs encode distinct proteins.

E Stanley1, S Ralph, S McEwen

  • 1Melbourne Tumour Biology Branch, Ludwig Institute for Cancer Research, Parkville, Victoria, Australia.

Molecular and Cellular Biology
|July 1, 1991
PubMed
Summary

Researchers identified two distinct forms of the lyn protein, a 56 kDa and a 53 kDa variant, arising from alternative splicing of a single gene. Both lyn protein isoforms are membrane-associated, suggesting a role in cell surface receptor signaling.

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • The lyn protein, a member of the src-related tyrosine kinase family, exists in different molecular weight forms.
  • Previous observations indicated the presence of 56 kDa and 53 kDa lyn proteins in various cell lines and tissues.

Purpose of the Study:

  • To clone and characterize the murine lyn cDNAs responsible for the observed protein isoforms.
  • To investigate the genetic basis and expression of these lyn protein variants.

Main Methods:

  • Cloning and sequencing of murine lyn cDNAs from an FDC-P1 cDNA library.
  • RNase protection analysis to detect corresponding mRNAs.
  • Sequence analysis of genomic clones to determine gene structure.
  • Expression of cDNAs in COS cells and protein analysis (immunoprecipitation, Western immunoblotting).

Related Experiment Videos

  • Subcellular fractionation to determine protein localization.

    • Main Results:

    • Two distinct murine lyn cDNAs, lyn11 (56 kDa) and lyn12 (53 kDa), were cloned and sequenced.
    • The 56 kDa lyn protein shares 96% similarity with human lyn.
    • The two proteins differ by a 21-amino-acid sequence.
    • Both lyn mRNAs are found in murine cells and tissues.
    • Alternative splicing of a single lyn gene generates these distinct mRNAs.
    • Expression in COS cells produced lyn proteins matching endogenous forms.
    • Both lyn isoforms are associated with the cell membrane.

    Conclusions:

    • A single lyn gene produces two distinct mRNA transcripts through alternative splicing, encoding 56 kDa and 53 kDa lyn proteins.
    • Both lyn protein isoforms are membrane-associated, supporting their potential role in signaling pathways involving cell surface receptors.