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Tate, P, Lee, M, Tweedie, S, Skarnes, WC and Bickmore, WA (1998) Capturing novel mouse genes encoding chromosomal and other nuclear proteins. J. Cell. Sci. 111 ( Pt 17):2575-85
The burgeoning wealth of gene sequences contrasts with our ignorance of gene function. One route to assigning function is by determining the sub-cellular location of proteins. We describe the identification of mouse genes encoding proteins that are confined to nuclear compartments by splicing endogeneous gene sequences to a promoterless betageo reporter, using a gene trap approach. Mouse ES (embryonic stem) cell lines were identified that express betageo fusions located within sub-nuclear compartments, including chromosomes, the nucleolus and foci containing splicing factors. The sequences of 11 trapped genes were ascertained, and characterisation of endogenous protein distribution in two cases confirmed the validity of the approach. Three novel proteins concentrated within distinct chromosomal domains were identified, one of which appears to be a serine/threonine kinase. The sequence of a gene whose product co-localises with splicesome components suggests that this protein may be an E3 ubiquitin-protein ligase. The majority of the other genes isolated represent novel genes. This approach is shown to be a powerful tool for identifying genes encoding novel proteins with specific sub-nuclear localisations and exposes our ignorance of the protein composition of the nucleus. Motifs in two of the isolated genes suggest new links between cellular regulatory mechanisms (ubiquitination and phosphorylation) and mRNA splicing and chromosome structure/function.
Amino Acid Sequence; Animals; Cell Line; Chromosomal Proteins, Non-Histone/chemistry; Chromosomal Proteins, Non-Histone/genetics; Embryo, Mammalian; Genes/genetics; Genetic Techniques; Ligases/genetics; Male; Mice; Molecular Sequence Data; Nuclear Envelope/genetics; Nuclear Proteins/chemistry; Nuclear Proteins/genetics; Phosphoprotein Phosphatases/chemistry; Phosphoprotein Phosphatases/genetics; Protein-Serine-Threonine Kinases/genetics; RNA Splicing; Ribonucleoprotein, U4-U6 Small Nuclear/genetics; Stem Cells; Ubiquitin-Protein Ligases
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