SPECIFIC, NONPRODUCTIVE CLEAVAGE OF PACKAGED BACTERIOPHAGE-T7 DNA IN-VIVO

The morphogenesis of bacteriophage T7 includes assembly of a procapsid that subsequently both packages DNA and changes in structure. The DNA packaged by T7 is concatemeric and is cleaved to mature length during packaging. In the present study, packaged DNA obtained from T7-iniected cells was analyzed after release from DNase-treated capsids. After fractionation by agarose gel electrophoresis, in-gel probing with oligonucleotides reveals that some of this DNA is shorter than mature T7 DNA; most of this short DNA has the T7 right end, but not the left end. Some of this short, packaged DNA is the product of left-to-right injection of DNA at the beginning of a T7 infection. However, subsequently produced short, packaged DNA has characteristics of a DNA that was produced during DNA packaging (incompletely packaged DNA or ipDNA). In contrast to results previously obtained in vitro, the profile of right-end-containing ipDNA is sometimes dominated by discrete bands. Some of the band-forming right-end-containing ipDNA appears with the kinetics of an abortive end product of packaging; cleavage in vivo appears to have arrested DNA packaging in this case. Other band-forming right-end-containing ipDNA appears with kinetics that have some characteristics expected of a precursor to the mature DNA; cleavage appears to have occurred after arrest of packaging in this case. The finding here of both left-to-right injection and right-to-left packaging is the most direct demonstration of polarity for these events in vivo. (C) 1995 Academic Press, Inc.