Interactions and reactions of ferritin with DNA.
Ferritin, normally considered a cytoplasmic iron-storage protein, is also found in the nuclei of some cells. There is no current agreement about its function(s) in this environment. Proposals include DNA protection, provision of iron to nuclear enzymes, and regulation of transcription initiation, but evidence for these functions is scanty. We have shown previously that H-ferritin subunits can be cross-linked to chromosomal DNA in vivo (Thompson, K. J., Fried, M. G., Ye, Z., Boyer, P., and Connor, J. R. (2002) J. Cell Sci. 115, 2165-2177). Here we describe systematic analyses of DNA binding and the covalent stability of DNA in the presence of ferritins from several different sources. Our data show that the H-subunit of human ferritin binds DNA, whereas neither the L-subunit nor the ferroxidase-deficient 222-mutant of the H-subunit has detectable binding activity. DNA binding is without significant preference for base composition, sequence, or the nature of DNA ends. H- and L-ferritins and ferritins of mixed subunit composition stimulate the conversion of superhelical plasmid DNA to the relaxed form. The sensitivity of this conversion to glycerol suggests that DNA is nicked by a free radical mechanism. The rate of nicking correlates with the iron content of the ferritin and is strongly inhibited by chelators. Ferritin-dependent nicking is characterized by a kinetic lag that is not seen in control reactions containing free iron species. These results suggest that the release of iron from ferritin is an important part of the nicking mechanism. The potential role of ferritin as a protector of the genome is discussed in the context of these results.
Surguladze
Nodar
N
Departments of Neural and Behavioral Science and Biochemistry and Molecular Biology, the Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033, USA.
Thompson
Khristy M
KM
Beard
John L
JL
Connor
James R
JR
Fried
Michael G
MG
D002021:Buffers; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D004278:DNA, Superhelical; D004305:Dose-Response Relationship, Drug; D004587:Electrophoresis, Agar Gel; D005293:Ferritins / Q000737:chemistry*; D005609:Free Radicals; D006801:Humans; D007501:Iron / Q000737:chemistry; D007700:Kinetics; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D013997:Time Factors; D014018:Tissue Distribution
D002021:Buffers; D004278:DNA, Superhelical; D005609:Free Radicals; D011994:Recombinant Proteins; D004247:DNA; D005293:Ferritins; D007501:Iron