Reconstitution of Colicin E 2 -Induced Deoxyribonucleic Acid Degradation in Spheroplast Preparations

Spheroplasts are insensitive to colicin E(2) and do not show deoxyribonucleic acid (DNA) degradation even in the presence of massive amounts of E(2). However, when both endonuclease I and E(2) were present, spheroplast DNA was degraded by an endonucleolytic activity which gave rise primarily to double-strand DNA cleavages, producing fragments having an average molecular weight of 9 x 10(6). Pancreatic ribonuclease could substitute for colicin E(2) in the reconstitution system, but pancreatic deoxyribonuclease could not replace endonuclease I. However, colicin E(2) could not activate transfer ribonucleic acid-inhibited endonuclease I in an in vitro system where pancreatic ribonuclease caused full stimulation.