Trans and cis requirements for intron mobility in a prokaryotic system.
Intron mobility requires cleavage of an intronless allele by an intron-encoded endonuclease, followed by transfer of the intron into the cleaved recipient. The mobile phage introns provide an opportunity to identify accessory functions involved in the intron inheritance process. To test for trans and cis requirements of mobility in Escherichia coli, we have exploited the td intron of phage T4 in both phage T4 and lambda genetic backgrounds. Mobility depends on host or phage recombinase functions, RecA or UvsX, respectively. The process also requires a phage-encoded 5'----3' exonuclease activity and associated annealing function that can be provided by phage lambda. Finally, host-encoded 3'----5' exonuclease activities are also implicated in intron inheritance. We demonstrated further that restriction enzymes could substitute for the intron-encoded endonuclease, indicating that the endonuclease does not have an essential role in recombination. Neither the precise position nor the nature of the double-strand break was critical to intron transfer. These features provide insight into the recombination pathway and are factors impacting on the spread of introns throughout natural populations.
Clyman
J
J
Molecular Genetics Program, Wadsworth Center for Laboratories and Research, State University of New York, Albany 12201-0509.
Belfort
M
M
D000251:Adenosine Triphosphatases / Q000378:metabolism; D010582:Bacteriophage lambda / Q000235:genetics*; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics*; D005092:Exonucleases / Q000378:metabolism; D005785:Gene Conversion; D005816:Genetic Complementation Test; D007438:Introns; D008565:Membrane Proteins / Q000378:metabolism; D010957:Plasmids; D011938:Rec A Recombinases / Q000378:metabolism; D011995:Recombination, Genetic; D013604:T-Phages / Q000235:genetics*; D014764:Viral Proteins / Q000378:metabolism
D004268:DNA-Binding Proteins; D008565:Membrane Proteins; C092863:UvsX protein, Enterobacteria phage T4; D014764:Viral Proteins; D004247:DNA; D011938:Rec A Recombinases; D005092:Exonucleases; D000251:Adenosine Triphosphatases