Purification of 14C-labelled deoxyribonuclease II from HeLa S3 lysosomes and its use as a marker for the study of nuclear deoxyribonuclease II

Abstract

Deoxyribonuclease II (DNAase II) in mammalian cells has generally been considered to be located in the lysosomes. Several recent studies have indicated that some DNAase II activity is present in purified nuclei; this, however, could have been due to some contamination of the nuclear fraction by lysosomes, or alternatively, it could have been caused by specific binding of lysosomal DNAase II to the nuclear fraction during isolation. Our previous studies have eliminated the possibility that lysosomal contamination was the cause of the presence of DNAase II in isolated nuclei. In this study I have purified 14C-labelled lysosomal DNAase II and added it to cells during isolation of their nuclei. This study demonstrates that there is no specific binding of lysosomal DNAase II to the nuclear fraction and concludes that DNAase II activity observed in isolated nuclei represents an intrinsic activity that might be involved in nuclear DNA metabolism.