Identification of autonomous IAP LTR retrotransposons mobile in mammalian cells.
Mammalian genomes contain two main classes of retrotransposons, the well-characterized long and short interspersed nuclear elements, which account for approximately 30% of the genome, and the long terminal repeat (LTR) retrotransposons, which resemble the proviral integrated form of retroviruses, except for the absence of an envelope gene in some cases. Genetic studies confirmed mobility of the latter class of elements in mice, with a high proportion of phenotypic mutations consequent to transposition of the intracisternal A particle (IAP) family of LTR retrotransposons. Using the mouse genome sequence and an efficient ex vivo retrotransposition assay, we identified functional, master IAP copies that encode all the enzymatic and structural proteins necessary for their autonomous transposition in heterologous cells. By introducing mutations, we found that the three genes gag, prt and pol are all required for retrotransposition and identified the IAP gene products by electron microscopy in the form of intracellular A-type particles in the transfected cells. These prototypic elements, devoid of an envelope gene, are the first LTR retrotransposons autonomous for transposition to be identified in mammals. Their high rates of retrotransposition indicate that they are potent insertional mutagens that could serve as safe (noninfectious) genetic tools in a large panel of cells.
Dewannieux
Marie
M
Unité des Rétrovirus Endogènes et Eléments Rétroïdes des Eucaryotes Supérieurs, UMR8122 CNRS, Institut Gustave Roussy, 94805 Villejuif, France.
Dupressoir
Anne
A
Harper
Francis
F
Pierron
Gérard
G
Heidmann
Thierry
T
D000818:Animals; D002415:Cats; D017931:DNA Primers / Q000737:chemistry; D015683:Gene Products, gag / Q000235:genetics / Q000378:metabolism; D007426:Genes, Intracisternal A-Particle / Q000235:genetics*; D015751:Genes, pol / Q000502:physiology; D006367:HeLa Cells / Q000648:ultrastructure; D006801:Humans; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D010641:Phenotype; D010957:Plasmids; D020079:Terminal Repeat Sequences / Q000235:genetics*; D014162:Transfection
D017931:DNA Primers; D015683:Gene Products, gag; D008666:Metalloendopeptidases; C071300:Prt1 protease