Gene trap insertion reveals two open reading frames in the mouse SSeCKS gene: the form predominantly detected in the nervous system is suppressed by the insertion while the other, specific of the testis, remains expressed.
Scaffold proteins play an important role in regulating signal transduction by targeting kinases and phosphatases in close proximity to their relevant substrates. SSeCKS protein has been described as a protein kinase C and A (PKC/PKA) anchoring protein as well as a PKC substrate with a tumor suppressor activity. In this study, we report the generation, via gene trapping in embryonic stem cells of mice carrying an insertion in the mouse SSeCKS gene. Through the molecular analysis of the insertion site, we show that SSeCKS contains two alternative promoters directing the synthesis of mRNAs (P1- and P2-mRNA), encoding two different proteins, one of which would be a truncated form of the other. Interestingly, these RNAs are differentially expressed, P2 being found exclusively in the male germ line, while P1 exhibits a dynamic and wider pattern of expression during embryonic development and in the adult; its expression is predominant in the nervous system. Finally, we show that P1- but not P2-mRNA expression is abolished by the insertion and furthermore that mice homozygous for the mutation lack SSeCKS in all tissues except the male germ cells. Nevertheless and surprisingly, these mice do not exhibit any obvious phenotype. The functional implications of these observations are discussed.
Camus
A
A
Unité de Biologie du Développement, URA-CNRS 1960, Institut Pasteur, 25 rue du Dr Roux, 75724 Cedex 15, Paris, France.
Mesbah
K
K
Rallu
M
M
Babinet
C
C
Barra
J
J
D054758:A Kinase Anchor Proteins; D000367:Age Factors; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D018797:Cell Cycle Proteins; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004622:Embryo, Mammalian / Q000378:metabolism; D005697:Galactosides / Q000378:metabolism; D005821:Genetic Techniques; D005854:Germ Cells / Q000166:cytology; D006579:Heterozygote; D006720:Homozygote; D007211:Indoles / Q000378:metabolism; D008297:Male; D051379:Mice; D008934:Mitogens / Q000096:biosynthesis* / Q000235:genetics*; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D009420:Nervous System / Q000196:embryology*; D016366:Open Reading Frames; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013234:Stem Cells / Q000378:metabolism; D013737:Testis / Q000196:embryology*; D014018:Tissue Distribution; D001616:beta-Galactosidase / Q000378:metabolism
D054758:A Kinase Anchor Proteins; C491161:Akap12 protein, mouse; D018797:Cell Cycle Proteins; D018076:DNA, Complementary; D005697:Galactosides; D007211:Indoles; D008934:Mitogens; D012333:RNA, Messenger; D001616:beta-Galactosidase; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside