Study of the interaction of glyceraldehyde-3-phosphate dehydrogenase with DNA.
Glyceraldehyde-3-phosphate dehydrogenase binds to homologous and heterologous single-stranded but not double-stranded DNA. Binding to RNA, poly(A) and poly(dA-dT) has also been observed. Enzyme binding to these nucleic acids leads to the formation of an insoluble complex which can be sedimented at low speed. The interaction of glyceraldehyde-3-phosphate dehydrogenase with DNA is strongly inhibited by NAD and NADH but not by NADP. Adenine nucleotides, which inhibit the dehydrogenase activity by competing with NAD for its binding site (Yang, S.T. and Deal, W.C., Jr. (1969) Biochemistry 8, 2806--2813), also inhibit enzyme binding to DNA, whereas glyceraldehyde-3-phosphate and inorganic phosphate are non-inhibitory. These results suggest that DNA interacts through the NAD binding sites of glyceraldehyde-3-phosphate dehydrogenase. In accordance with this idea, it was found that DNA also binds to lactate dehydrogenase, an enzyme containing a similar dinucleotide binding domain, and that this binding is inhibited by NADH. A study of the base specificity of the DNA-glyceraldehyde-3-phosphate dehydrogenase interaction using dinucleoside monophosphates shows that inhibition of DNA binding by the dinucleotides requires the presence of a 3'-terminal adenosine and is greater when the 5'-terminus contains a pyrimidine instead of a purine. These results suggest that the dinucleotides bind at the NAD site of the dehydrogenase and that the enzyme would interact preferentially with PypA dinucleotides present in the nucleic acid.
D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D006224:Cricetinae; D004247:DNA / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism*; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008647:Mesocricetus; D009243:NAD / Q000494:pharmacology; D009249:NADP / Q000494:pharmacology; D009691:Nucleic Acid Denaturation; D009711:Nucleotides / Q000494:pharmacology; D009994:Osmolar Concentration; D011061:Poly A / Q000378:metabolism; D011067:Poly dA-dT / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D013696:Temperature
D009711:Nucleotides; D009243:NAD; D011061:Poly A; D011067:Poly dA-dT; D008345:Manganese; D009249:NADP; D012313:RNA; D004247:DNA; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D008274:Magnesium