Intron mobility in phage T4 is dependent upon a distinctive class of endonucleases and independent of DNA sequences encoding the intron core: mechanistic and evolutionary implications.
Although mobility of the phylogenetically widespread group I introns appears to be mechanistically similar, the phage T4 intron-encoded endonucleases that promote mobility of the td and sunY introns are different from their eukaryotic counterparts. Most notably, they cleave at a distance from the intron insertion sites. The td enzyme was shown to cleave 23-26 nt 5' and the sunY endonuclease 13-15 nt 3' to the intron insertion site to generate 3-nt or 2-nt 3'-OH extensions, respectively. The absolute coconversion of exon markers between the distant cleavage and insertion sites is consistent with the double-strand-break repair model for intron mobility. As a further critical test of the model we have demonstrated that the mobility event is independent of DNA sequences that encode the catalytic intron core structure. Thus, in derivatives in which the lacZ or kanR coding sequences replace the intron, these marker genes are efficiently inserted into intron-minus alleles when the cognate endonuclease is provided in trans. The process is therefore endonuclease-dependent, rather than dependent on the intron per se. These findings, which imply that the endonucleases rather than the introns themselves were the primordial mobile elements, are incorporated into a model for the evolution of mobile introns.
Bell-Pedersen
D
D
Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201-0509.
Quirk
S
S
Clyman
J
J
Belfort
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D005075:Biological Evolution; D004251:DNA Transposable Elements; D004279:DNA, Viral / Q000235:genetics*; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D004720:Endonucleases / Q000378:metabolism*; D007438:Introns; D008969:Molecular Sequence Data; D013604:T-Phages / Q000201:enzymology / Q000235:genetics*
D004251:DNA Transposable Elements; D004279:DNA, Viral; D004706:Endodeoxyribonucleases; D004720:Endonucleases; C065074:I-TEVI endonuclease; C065073:I-TEVII endonuclease