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Related Experiment Videos

Capturing novel mouse genes encoding chromosomal and other nuclear proteins

P Tate1, M Lee, S Tweedie

  • 1MRC Human Genetics Unit, Western General Hospital, Crewe Road, Edinburgh EH4 2XU, Scotland, UK.

Journal of Cell Science
|August 14, 1998
PubMed
Summary

Researchers identified mouse genes for proteins within specific nuclear compartments using a gene trap method. This reveals novel proteins and links ubiquitination and phosphorylation to mRNA splicing and chromosome function.

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Area of Science:

  • Cell Biology
  • Genetics
  • Molecular Biology

Background:

  • Vast gene sequences exist, but gene functions remain largely unknown.
  • Determining protein sub-cellular localization is a key strategy for assigning gene function.
  • The nucleus contains numerous compartments with poorly understood protein compositions.

Purpose of the Study:

  • To identify mouse genes encoding proteins localized to specific nuclear compartments.
  • To characterize novel nuclear proteins and their functions.
  • To explore potential links between identified proteins and cellular regulatory mechanisms.

Main Methods:

  • Utilized a gene trap approach by splicing endogenous gene sequences to a promoterless betageo reporter in mouse embryonic stem (ES) cells.
  • Screened ES cell lines for betageo fusions localized to sub-nuclear compartments like chromosomes, nucleolus, and splicing factor foci.

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  • Determined the sequences of 11 trapped genes and validated protein localization for two genes.

    • Main Results:

    • Successfully identified 11 trapped genes encoding proteins with specific sub-nuclear localizations.
    • Discovered three novel proteins localized to distinct chromosomal domains, including a putative serine/threonine kinase.
    • Identified a gene product co-localizing with spliceosome components, suggesting a role as an E3 ubiquitin-protein ligase.
    • The majority of isolated genes represent novel, uncharacterized genes.

    Conclusions:

    • The gene trap approach is effective for identifying genes with novel proteins and specific nuclear localizations.
    • The study highlights significant gaps in our understanding of the nucleus's protein content.
    • Identified protein motifs suggest new connections between ubiquitination, phosphorylation, mRNA splicing, and chromosome biology.