Characterization of the mIF4G Domains in the RNA Surveillance Protein Upf2p

Round 1

Reviewer 1 Report

In the manuscript entitled, “Characterization of the mIF4G domains in the RNA surveillance protein Upf2p”, the authors characterized Upf2p protein and found a region in the Upf2p’s domains that is important for NMD activity. Overall, the manuscript is comprehensive, and the contents of the manuscript are very well portrait. This manuscript should be accepted with minor editing in the CIMB journal.

Comments

1. Line #91; The yeast 2 μ plasmid pG-1.., please correct the value.

2. In the alignment of figure 4; only 8 sites have been indicated for phosphorylation, while you have found 12 phosphorylation sites; please indicate all the phosphorylation sites in this figure.

3. Figure 6; “SC-Trp-Arg plates containing either 0, 200 or 100 micro-g of….”’,In the figure however, 250 micro-g plate has been mentioned, please add the corresponding text to the legend/manuscript. Also in Figure 8, please correct the unit of canavanine.

4. Have you studied the structure of mIF4G I domain, which was reported previously, to see the location of D59. Please write one or two sentences about the location of D59 in the manuscript and explain whether deletion would change the structure or not.

5. Discussion section is little big; please consider removing some non-essential part(s) from this section.

Minor English editing required.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

The manuscript communicates an interesting study on a relevant subject that has attracted attention recently in the biology of the eukaryotic cell, nonsense-mediated mRNA decay. The results are sound and support well the main conclusions. I have only two points to offer for manuscript strengthing.

The methodology is mostly cryptic, and more details about the experimental conditions should be included. As it is, it seems not possible to replicate the study.

The choice for amino acid substitutions in site-directed mutagenesis seems controversial, the change of a negatively charged amino acid for a positive one, and vice versa, may alter the structure of the domain. I think the authors should take with caution the associated results and if possible, should include evidence that no major structural changes are expected with those changes.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

My concerns were addressed, thank you.