H1 histone modulates DNA hydrolysis with WEN1 and WEN2 endonucleases from wheat coleoptiles.
We show that total H1 histone from wheat seedlings or rat liver enhances hydrolysis of lambda phage DNA with plant endonucleases WEN1 and WEN2 isolated from wheat coleoptiles. Optimal DNA/protein weight ratio in the hydrolysis reaction is 1 : 1. The action of fractions I and IV (obtained from total wheat H1 histone by electrophoresis) on DNA hydrolysis with WEN1 and WEN2 enzymes depends on the DNA methylation status. Fraction IV of wheat histone H1 stimulates hydrolysis of unmethylated lambda phage DNA with WEN1 and WEN2 enzymes. Hydrolysis of methylated lambda phage DNA (it contains 5-methylcytosine in Cm(5)CWGG sequences and N(6)-methyladenine in Gm(6)ATC sites) with WEN1 is inhibited with fractions I and IV of wheat H1 histone. Fractions II and III of wheat H1 histone do not influence DNA hydrolysis with WEN1 and WEN2. S-Adenosyl-L-methionine (SAM) stimulates activity of these plant enzymes. But in the presence of H1 histone, SAM does not add to the ability of the enzyme to hydrolyze more DNA compared with that induced with H1 histone itself. Therefore, the stimulating effects of SAM and H1 histone on DNA hydrolysis with plant endonucleases may be similar. It could be suggested that SAM and H1 histone can induce more or less analogous allosteric transformations in the structure of the investigated plant endonucleases. Thus, DNA hydrolysis with plant endonucleases is modulated with total H1 histone. H1 histone fractions affect DNA hydrolysis in a different fashion; they enhance or inhibit hydrolysis depending on the DNA methylation status. We suggest that H1 histone changes site specificity of endonucleases or it might be responsible for formation of new or masking of old sites available for these enzymes due to changes in DNA structure induced in a DNA-histone complex.
Fedoreyeva
L I
LI
Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Russia.
Smirnova
T A
TA
Kolomijtseva
G Ya
GY
Vanyushin
B F
BF
D000818:Animals; D001435:Bacteriophages / Q000378:metabolism; D018548:Cotyledon / Q000378:metabolism*; D019175:DNA Methylation; D004279:DNA, Viral / Q000737:chemistry*; D004720:Endonucleases / Q000737:chemistry* / Q000302:isolation & purification; D006657:Histones / Q000737:chemistry* / Q000302:isolation & purification; D006868:Hydrolysis; D008099:Liver / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification; D051381:Rats; D014908:Triticum / Q000378:metabolism*
D004279:DNA, Viral; D006657:Histones; D010940:Plant Proteins; D004720:Endonucleases