Enhanced translation of mRNAs encoding proteins involved in mRNA translation during recovery from heat shock
Authors: Jnanankur Bag
Andrea-Kaye Datu
Journal: PLoS ONE (2013)
Discipline: Multidisciplinary
Keywords: mRNA; translation; PABP1; eEF1A; RPS6
Affiliations: University of Guelph, Department of Molecular & Cellular Biology, Guelph, Ontario, Canada.

Abstract:  The mRNAs encoding poly (A) binding protein (PABP1), eukaryotic elongation factor 1A (eEF1A) and ribosomal protein S6 (RPS6) belong to the family of terminal oligo pyrimidine tract (TOP) containing mRNAs. Translation of the TOP mRNAs is regulated by growth signals and usually codes for proteins involved in mRNA translation. Previous studies from our laboratory showed that translation of PABP1 mRNA was preferentially enhanced during recovery of HeLa cells from heat shock. Presence of the 5' TOP cis element was required for the observed increase of PABP1 mRNA translation. In the studies reported here we showed that translation of two additional TOP mRNAs such as, eEF1A and RPS6 was similarly enhanced during recovery. In addition, we showed by in vivo cross-linking experiments that the cellular nucleic acid binding protein ZNF9 binds to all three TOP mRNAs examined in these studies as well as to the β-actin mRNA that lacks a TOP cis element. Binding of ZNF9 to mRNAs was observed in both heat-shocked and non heat- shocked cells. However, depletion of ZNF9 by siRNA prevented the preferred stimulation of PABP1, eEF1A and RPS6 expression during recovery from heat shock. There was no detectable effect of ZNF9 depletion on the basal level of expression of either β-actin or PABP1, eEF1A and RPS6 in HeLa cells following recovery from heat shock. Conclusion: Although the presence of ZNF9 was required for the translational stimulation of PABP1, eEF1A and RPS6 mRNAs, the mechanistic details of this process are still unclear. Since ZNF9 was shown to bind both TOP and non-TOP mRNAs, it is uncertain whether ZNF9 exerts its stimulatory effect on TOP mRNA translation following recovery from heat shock through the TOP cis-element. Perhaps additional factors or post-translational modification(s) of ZNF9 following heat shock are necessary for the preferred increase of TOP mRNA translation. 

Retracted: Yes

PubPeer comments:
null, 
 Reposting the concerns about Figure 3A, which did not survive the PubPeer upgrade. ![file] , 
 2021 retraction. https: journals.plos.org plosone article?id=10.1371 journal.pone.0246558 Following the publication of this article [1], concerns were raised regarding results presented in Figs 1, 2, 3, 4, and 5. Specifically, In Fig 1B, vertical discontinuities suggestive of image splicing were detected on either side of the 5 u00b5L lane of the PABP1 panel. The PARP1 panel and the eEF1A2 panel in Fig 2A appear similar. The authors indicated that the wrong panel was used for the eEF1A2 results. The C (-RT) panel in Fig 2B appears to be devoid of any signal, including background signal usually expected in (negative) PCR results. In Fig 3A, vertical discontinuities suggestive of image splicing were detected between the first and second lane of both the PABP 1 C and PABP 1 R panels. In Fig 3A, the eEF1A1 HS panel appears similar to the eEF1A2 R panel when flipped, the eEF1A1 R panel appears similar to the eEF1A2 C panel when flipped, and the first and second lane of the eEF1A1 C panel appear similar to the second and third lane of the eEF1A2 HS panel when flipped. For the eEF1A1 C, eEF1A1R, eEF1A2 HS and eEF1A2 C gels, different quantification numbers are reported for bands that appear similar. The authors indicated that the wrong panels were used for the eEF1A2 results. In Fig 5A, vertical discontinuities suggestive of image splicing were detected between the second and third lane of the ZNF9 panel. The quantification results presented for the protein abundance in Figs 1C and 5C, the relative mRNA levels in Figs 2C and 4B, and the % of total mRNA results in Fig 3B do not always appear to match the blot and gel data presented in the related figures. In addition, there appears to be less variation in the results and the error bars than would be expected. The authors have provided photographs of lab book results underlying some of the published figures. However, the results provided did not include any markers or other control bands to confirm the results. In addition, some of the results presented cropped images of PCR and blot results only, as opposed to the full, uncropped figures underlying the published images. In light of the concerns affecting multiple figure panels that question the integrity of these data and the data handling by the research group, the PLOS ONE Editors retract this article. JB did not agree with the retraction and stands by the articleu2019s findings. AKD either did not respond directly or could not be reached. Reference 1.Datu A-K, Bag J (2013) Enhanced Translation of mRNAs Encoding Proteins Involved in mRNA Translation during Recovery from Heat Shock. PLoS ONE 8(5): e64171. https: doi.org 10.1371 journal.pone.0064171 pmid:23696868 ,