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closeA comment about the authenticity of the paper
Posted by wushuai on 24 Nov 2011 at 08:38 GMT
In this paper，the author doesn't provide any evidences that MYCT1 and MYCT1-TV are the same thing in vivo. What I mean is one of them is not exist in vivo. Because they has no data to detect the protein level but only the mRNA level. The question is the mRNAs of MYCT1 and MYCT1-TV is almost the same except the N terminal. So, the primers used in this paper to detect the two mRNAs can not identify each of them. Because each mRNA can confuse the detection of the mRNA. So, it is no doubt that all the experiments in this paper to detect the mRNAs level of MYCT1 and MYCT1-TV have no difference. Maybe in the paper, the author use different qPCR primers to detect the same mRNA in all their experiments. I think the author should give more data to prove MYCT1 and MYCT1-TV are different thing and they all are not artifical CDSs in vivo.
RE: A comment about the authenticity of the paper
fuweineng replied to wushuai on 29 Nov 2011 at 06:41 GMT
Thank you for your attention on our work. It is a real good question. In the study, we aimed to confirm the transcriptional and translational start sites of MYCT1 predicted by our group in the previous study through corresponding experiments. During study, we discovered two transcripts and the primers used in this paper can identify each of them, suggesting that it harbored a new transcript of MYCT1. Although the protein sequences the two transcripts encode for are similar, whether they are the same proteins or not needs to be further studied. We are currently seeking an appropriate way to solve this problem. The preparation of anti-MYCT1-TV and anti- MYCT1 antibodies is on the way, so we can only prove the expression and existing of two transcripts from mRNA level at presence. We will be grateful if you kindly give us some good advice on it? Thanks again.
RE: RE: A comment about the authenticity of the paper
Klaus_Roemer replied to fuweineng on 12 Jan 2012 at 06:34 GMT
RACE and cDNA cloning, when carefully performed and controlled, are established procedures for the identification of novel transcript variants. The alignment of MYCT1-TV (the variant) and MYCT1 (Figure 2) suggests differences in the 5’ and 3’ regions, as well as, importantly, predicted protein sequence differences between the N-termini. Even if the ‘two’ predicted proteins will eventually turn out to be ‘identical’ (in primary sequence and/or function), the variations in the mRNA (cDNA) sequences alone – and in particular those concerning the 3’ region – may already be of great importance and interest as they may add further degrees of freedom for regulation. For example, 3’ flanking regions are frequently subject to regulation by microRNAs. If discriminating antibodies will permit the identification of two distinct proteins in future work, this will of course be of great help; however, I don’t see necessity for this to be part of the present study. Even if, in the end, MYCT1/-TV should turn out to be identical in function and regulation, there is plenty of new data and insight in this paper that goes well beyond the distinction of MYCT1 and MYCT1-TV.
RE: RE: RE: A comment about the authenticity of the paper
wushuai replied to Klaus_Roemer on 20 Mar 2012 at 16:39 GMT
Thank you for your concern. I think your opinion is farsighted. It is very likely that 5'-UTR and 3'-UTR participate into the regulation of mRNA. 3’ flanking regions are frequently subject to regulation by microRNAs. But, even if the two transcripts have different 3-UTR(MYCT1-TV has longer 3'-UTR) , in this paper, the expression levels of MYCT1 an TV are equal in vitro and in vivo. so, it seems that 3-UTR not affect the TV mRNA level. So, even if there exists two transcripts, whether it exists two proteins as pridicted. In addition, the two transcripts have almost same N-terminal and different C-terminal，but the ORFs are same. Does the translation machine can identify which AUG is the translation origin. And we also cannot determine which transcript translates into one of the "'two proteins".and whether one of the "two proteins" is artificial. So, It is urgent to prove whether the "two proteins"are identical or truely two different one.
RE: RE: RE: A comment about the authenticity of the paper
wushuai replied to Klaus_Roemer on 20 Mar 2012 at 16:39 GMT
Thank you for your reply. first, I want to apologize for my careless. The primers used in your paper can identify the two transcripts. But, as you say,we have no discriminating antibodies to identify "the two proteins" The two transcripts have almost same N-terminal and different C-terminal，but the ORFs are same. So, even if it exists two transcripts, I think it is impossible to identify whether MYCT1 mRNA exoress protein MYCT1 or MYCT1-TV express protein MYCT1 and vice versa. What I mean is the translation machine cannot identify whether the first AUG or second is the real translational origin site. In general, the ribosome will choose the first AUG the encounter as translation origin, so it is weird MYCT1-TV can translate into the protein predicted. If this is true, it is suggested that there exists a mechism to determine which AUG is origin site. More work should to focus on whether the "two" proteins are the same or one of them is not exist.
RE: RE: A comment about the authenticity of the paper
wushuai replied to fuweineng on 20 Mar 2012 at 16:40 GMT
Thank you for your reply. first, I want to apologize for my careless. The primers used in your paper can identify the two transcripts. But, as you say,we have no discriminating antibodies to identify "the two proteins" The two transcripts have almost same N-terminal and different C-terminal，but the ORFs are same. So, even if it exists two transcripts, I think it is impossible to identify whether MYCT1 mRNA exoress protein MYCT1 or MYCT1-TV express protein MYCT1 and vice versa. What I mean is the translation machine cannot identify whether the first AUG or second is the real translational origin site. In general, the ribosome will choose the first AUG the encounter as translation origin, so it is weird MYCT1-TV can translate into the protein predicted. If this is true, it is suggested that there exists a mechism to determine which AUG is origin site. More work should to focus on whether the "two" proteins are the same or one of them is not exist.
RE: A comment about the authenticity of the paper
wushuai replied to wushuai on 20 Mar 2012 at 16:03 GMT
Thank you for your reply. first, I want to apologize for my careless. The primers used in your paper can identify the two transcripts. But, as you say,we have no discriminating antibodies to identify "the two proteins" The two transcripts have almost same N-terminal and different C-terminal，but the ORFs are same. So, even if it exists two transcripts, I think it is impossible to identify whether MYCT1 mRNA exoress protein MYCT1 or MYCT1-TV express protein MYCT1 and vice versa. What I mean is the translation machine cannot identify whether the first AUG or second is the real translational origin site. In general, the ribosome will choose the first AUG the encounter as translation origin, so it is weird MYCT1-TV can translate into the protein predicted. If this is true, it is suggested that there exists a mechism to determine which AUG is origin site. More work should to focus on whether the "two" proteins are the same or one of them is not exist.