Active participation of cellular chaperone Hsp90 in regulating the function of rotavirus nonstructural protein 3 (NSP3)
Authors: Dipanjan Dutta
Shiladitya Chattopadhyay
Koki Taniguchi
Mamta Chawla-Sarkar
Parikshit Bagchi
Anupam Mukherjee
Umesh Chandra Halder
Nobumichi Kobayashi
Satabdi Nandi
Journal: Journal of Biological Chemistry (2011)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: Heat shock protein 90; Hsp90; rotavirus replication; NSP3; 17-N,N-dimethylethylenediamine-geldanamycin
Affiliations: Division of Virology, National Institute of Cholera and Enteric Diseases, Beliaghata, Kolkata, India.

Abstract:  Heat shock protein 90 (Hsp90) has been reported to positively regulate rotavirus replication by modulating virus induced PI3K/Akt and NFŒ∫B activation. Here, we report the active association of Hsp90 in the folding and stabilization of rotavirus nonstructural protein 3 (NSP3). In pCD-NSP3-transfected cells, treatment with Hsp90 inhibitor (17-N,N-dimethylethylenediamine-geldanamycin (17DMAG)) resulted in the proteasomal degradation of NSP3. Sequence analysis and deletion mutations revealed that the region spanning amino acids 225-258 within the C-terminal eIF4G-binding domain of NSP3 is a putative Hsp90 binding region. Co-immunoprecipitation and mammalian two-hybrid experiments revealed direct interaction of the C-terminal 12-kDa domain of Hsp90 (C90) with residues 225-258 of NSP3. NSP3-Hsp90 interaction is important for the formation of functionally active mature NSP3, because full-length NSP3 in the presence of the Hsp90 inhibitor or NSP3 lacking the amino acid 225-258 region did not show NSP3 dimers following in vitro coupled transcription-translation followed by chase. Disruption of residues 225-258 within NSP3 also resulted in poor RNA binding and eIF4G binding activity. In addition, inhibition of Hsp90 by 17DMAG resulted in reduced nuclear translocation of poly(A)-binding protein and translation of viral proteins. These results highlight the crucial role of Hsp90 chaperone in the regulation of assembly and functionality of a viral protein during the virus replication and propagation in host cells. 

Retracted: No

PubPeer comments:
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 How to explain the repetitive elements in figures 5B, 6B and 7A? , 
 I agree with Drusus, in particular with Figure 7A. The longer one looks, the more features appear to be visible multiple times, either between the two panels, or within the same photo. Here is what I see (contrast changed to bring out more detail):  , 
 #1 Thank you for bringing this to our notice. we will make sure to submit the raw data to the journal. Authors , 
 #5 Setting aside all the repeated motifs, here is another way of looking at the similarity between the two panels of Fig 7A.  , 
 #1  , 
 Dear Hoya, as per the response posted couple of days ago by the first author, we are in process of corresponding with the journal to clarify the raised concerns. , 
 2019 Expression of Concern. http: www.jbc.org content 294 41 15118.short The publisher of the Journal of Biological Chemistry is issuing an Expression of Concern to inform readers that credible concerns have been raised regarding some of the data and conclusions in the article listed above. The Journal of Biological Chemistry will provide additional information as it becomes available. , 
 August September 2020 withdrawal: https: www.jbc.org content 295 39 13696.short * This article has been withdrawn by the authors. The Journal determined that there are similar repetitive features in the background area of Figs. 4B, 5B, 6B, and 7A and Fig. S4, but the authors do not agree to the raised concerns. The authors state that they provided uncropped scans of the autoradiograms from the corresponding and or replicate experiments done at same time of the original experiments. The authors state that due to the tropical climate, the quality of the autoradiograms may have partially deteriorated as the work was done more than 10 years ago, but still they could not visualize any repetitive elements in the background. However, the authors could not provide the original, uncropped electronic copies of the scans that were used for preparation of the published images. The authors state that all available hard copy autoradiograms present in the laboratory records were reviewed and verified favorably by the institutional review committee and a third-party forensic laboratory. The authors state that they had provided the cropped tiff files from the final assembled figures used during manuscript submission as well as figures downloaded from the JBC website to the forensic laboratory. Copies of both of the reports were shared with the Journal. The authors affirm that all the experiments were performed appropriately and the concerns raised do not affect the accuracy of the results or the conclusions of the scientific work. However, in the interest of maintaining the accuracy in the published scientific literature, the authors wish to withdraw this article but plan to publish a new version of the article corroborating the findings of this work in the future. * , 
 https: retractionwatch.com 2020 09 30 author-says-misguided-efforts-for-the-ideal-western-blot-led-to-the-withdrawal-of-these-studies ,