8 replies to this topic

[cardosopedro]

Hi.

I am studying a protein which is probably a transcription factor (it encodes a transcription factor domain although this domain is not involved in DNA-binding). Bioinformatics predictions show that apparently the protein does not have DNA-binding motifs, although I have functional data highly suggestive of the protein being involved in gene expression regulation.

Is it possible that it is a non-binding transcription factor? Are these proteins common? Do they have particular features? Could it be interacting with a DNA-binding molecule allowing the regulation of gene expression? Does anyone know how I can predict this and look for the putative mediator(s)?


A big thanks for anyone who can help!

Edited by cardosopedro, 14 January 2011 - 08:09 AM.

[cardosopedro]

Can someone help me please?

[bob1]

Well there are the E2F group and Rb that don't bind to DNA but do control transcription at a certain level. You could do ChIP to identify if it is binding DNA and which sequence it is binding to.

[cardosopedro]

Thank you.

Yes, I thought of that. But there is not comercially available antibody against my protein. I will have to produce my own antibody.

[Radish]

I would proppose that you cloned your protein from total RNA, add a tag (flag or myc) get into a vector (gateway system would probably be the best.
And in 2 to 3 months you could overexpress it in your target cells and use a flag or myc anti-body to do the pull-down.


best regards

Radish

[cardosopedro]

Very nice idea, thanks!

[Radish]

No problem.
Pm me if you need any help!

[gaurav amit prakash]

hey

as we all know that there are two type of tfs ;
1) gtf
2) taf
so first of all u have to see that they are working as gtfs or tafs
for gtfs u have to use chip to see that either they are binding to dna or not

for tafs u have to see it by south western blotting.

check it it may say something.....

[Radish]

This may sound stupid but...
what are tafs and gtfs?


((TBP)-associated factors and General transcription factors??)

if so you don't have limited techniques for any particular class...