With treatment of cell lines ( cardiofibroblast cells), changes in mRNA level of the gene of interest has been observed overtly, but there is no change in protein level. I am wondering how this can be explained...
Hi Fereshteh,
how do you measure the changes in mRNA and protein level and how big is the change in mRNA level?
There are many ways how expression and protein synthesis is regulated maybe some kind of post-transcriptional regulation is happening? Post-transcriptional regulation might influence translation initiation and/or mRNA stability. Often regulatory RNAs play a role.
Maybe you should try other timepoints for your protein levels. Do you know something about the regulation of your goi?
all the best
Ulrike
Hi Fereshteh,
how do you measure the changes in mRNA and protein level and how big is the change in mRNA level?
There are many ways how expression and protein synthesis is regulated maybe some kind of post-transcriptional regulation is happening? Post-transcriptional regulation might influence translation initiation and/or mRNA stability. Often regulatory RNAs play a role.
Maybe you should try other timepoints for your protein levels. Do you know something about the regulation of your goi?
all the best
Ulrike
Could you be looking at alternative splicing ?
Yes , post translation modification could also be a factor.
Dear Ulrike,
thank you for the response. Yes , the difference between mRNA level is tangible. Unfortunately, the regulation of protein expression is under investigation. I was also thinking about miRNA. but this also can affect not only protein level but also protein level.
Regards,
Fereshteh
Hi Fereshteh,
in human tissues, I have measured many different mRNA and proteins are rarely saw correlations between them. mRNA are often less stable than proteins and are more susceptible to degradation. If your proteins of interest are secreted, you might consider measuring them in cell media. The method of extraction can also affect your results ex. whole cell extract vs specific subcellular compartments.
Sincerely,
Cynthia Picard, Ph.D.
Hi
The half life of the mRNAs and the corresponding protein vary depending on the types or you can say their mechanism and rate of degradation is diverse. If the half life of the protein is greater/less than that of their corresponding mRNA inverse relationship you will observe.
The most likely explanation is that the mRNA is not recruited into polyribosomes and thus it cannot be translated. I recommend you isolate polysomes from the tissue in questions and measure the mRNA in a) the total extract, b) in the polysome fraction, c) in the non-polysome fraction. You can find many polysome isolation techniques in my profile or even read "Where has all the message gone?" Best of luck. Eric Davies
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