Difference between histone's and DNA's methyltransferase?

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Difference between histone's and DNA's methyltransferase?

Post by character » Mon Dec 28, 2009 9:18 am

I want to know the typical differences between histone methyltransferase and DNA transferase . How could I detect enzyme activity of histone methyltransferases in vitro? Anything you can tell me would me much appreciated, thanks

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Post by kolean » Mon Dec 28, 2009 9:33 pm

Detection of methylated histone tails is usually done with antibodies to the specific methylation that you are studying (even mono-, di-, or tri-methylation of lysines at specific histone- H3, H4, H1, etc.).

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Post by MrMistery » Sat Jan 02, 2010 5:59 pm

well generally when you want to measure activity of an enzyme in vitro you need to design an assay to measure it. So you will need to have a substrate there ( a histone tail fragment) and a way to detect the methylation (no idea how). One problem i don't think you would have is detecting between histone and dna methyltransferases, since you wouldn't have any DNA present in your assay
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Re: Difference between histone's and DNA's methyltransferase?

Post by zami'87. » Mon Jan 18, 2010 10:10 am

I dunno if it helps..but you can detect DNA methylation by methylation specific PCR (MSP).
First you treat DNA with sodium bisulfite under alkaline conditions.Result is conversion of unmethylated Cs to Us.Methylated Cs are unchanged.Then add specific primer that will anneal only if that segment of DNA is methylated(so C is not converted to U).If primer anneals and you get reaction then C was methylated..Use it to measure activity of enzyme somehow :s.
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