Polynucleotide:adenosine glycosidase activity of saporin-L1: effect on various forms of mammalian DNA.
Saporin-L1 from the leaves of Saponaria officinalis belongs to a group of plant polynucleotide:adenosine glycosidases, known as ribosome-inactivating proteins due to their property of depurinating the major rRNA. Previous experiments indicated that saporin-L1 and other ribosome-inactivating proteins depurinate also DNA [Barbieri et al. (1994) Nature 372, 324; and (1996) Biochem. J. 319, 507-513]. Here we describe the effects of highly purified nuclease-free saporin-L1 on mammalian nuclear and mitochondrial DNA. Saporin-L1 had less activity on mitochondrial DNA than on nuclear DNA. A low, although significant, depurination of both chromatin and whole nuclei was observed. Mitochondrial nucleic acids are heavily depurinated in intact mitochondria, although the contribute of mtDNA to the deadenylation events is not known. The kinetic constants for several substrates were determined.
Barbieri
L
L
Dipartimento di Patologia Sperimentale, Università di Bologna, Via San Giacomo, 14, I-40126, Bologna, Italy. barbieri@alma.unibo.it
Valbonesi
P
P
Govoni
M
M
Pession
A
A
Stirpe
F
F
D000818:Animals; D004247:DNA / Q000378:metabolism*; D006801:Humans; D000922:Immunotoxins; D007700:Kinetics; D009699:N-Glycosyl Hydrolases / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism*; D051381:Rats; D054788:Ribosome Inactivating Proteins; D054789:Ribosome Inactivating Proteins, Type 1; D000076985:Saporins; D013379:Substrate Specificity
D000922:Immunotoxins; D010940:Plant Proteins; D054789:Ribosome Inactivating Proteins, Type 1; D004247:DNA; D009699:N-Glycosyl Hydrolases; D054788:Ribosome Inactivating Proteins; D000076985:Saporins