Arabidopsis Argonaute 2 regulates innate immunity via miRNA393(∗)-mediated silencing of a Golgi-localized SNARE gene, MEMB12
Authors: Hsien-Da Huang
Natasha Raikhel
Xiaoming Zhang
Surekha Katiyar-Agarwal
Shang Gao
Hongwei Zhao
Hailing Jin
Wei-Chi Wang
Journal: Molecular Cell (2011)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: Argonaute proteins; RNA silencing pathways; Arabidopsis AGO2; bacterial pathogen; Pseudomonas syringae pv. tomato
Affiliations: Department of Plant Pathology and Microbiology, University of California, Riverside, Riverside, CA 92521, USA.

Abstract:  Argonaute (AGO) proteins are critical components of RNA silencing pathways that bind small RNAs and mediate gene silencing at their target sites. We found that Arabidopsis AGO2 is highly induced by the bacterial pathogen Pseudomonas syringae pv. tomato (Pst). Further genetic analysis demonstrated that AGO2 functions in antibacterial immunity. One abundant species of AGO2-bound small RNA is miR393b(∗), which targets a Golgi-localized SNARE gene, MEMB12. Pst infection downregulates MEMB12 in a miR393b(∗)-dependent manner. Loss of function of MEMB12, but not SYP61, another intracellular SNARE, leads to increased exocytosis of an antimicrobial pathogenesis-related protein, PR1. Overexpression of miR393b(∗) resembles memb12 mutant in resistance responses. Thus, AGO2 functions in antibacterial immunity by binding miR393b(∗) to modulate exocytosis of antimicrobial PR proteins via MEMB12. Since miR393 also contributes to antibacterial responses, miR393(∗)/miR393 represent an example of a miRNA(∗)/miRNA pair that functions in immunity through two distinct AGOs: miR393(∗) through AGO2 and miR393 through AGO1. 

Retracted: No

PubPeer comments:
 Figure 2B: same blank panel was reused several times (red boxes), which casts doubts on whether these experiments were done for real. Could the authors please comment? ![file] , 
 Hi, thank you very much for identifying this terrible mistake, and I take this very seriously. I was so shocked to see the duplication because I have been always very careful to double check all the original results and blots for publication. I am so sorry that I was not careful enough at checking the blank negative blots this time, and I did not spot the big mistake of duplicated blank panels. After I saw your message, I immediately contacted the two First Authors, Xiaoming Zhang and Hongwei Zhao, to find the original blot images and to figure out how this mistake could have occurred. Luckily, Hongwei Zhao, my former postdoc who did all the AGO-IP experiments related to this figure, admitted that he made this terrible mistake, and managed to find the original U6 Northern blots (two biological replicates) and the original Western blot for the Tubulin control. All of these blots x-ray films were scanned in early 2011, and Hongwei also sent me the screenshots of these files to show their originated dates, which was in early 2011. I attached all these figures here. Hongwei Zhao explained to me why he made such a big mistake. He said during that time, he was extremely stressed out by his daughteru2019s upcoming heart surgery. He was distracted with all the doctoru2019s examinations and appointments for his daughteru2019s medical condition, and had to be off work regularly during that period of time. So he asked Xiaoming to help him develop the x-ray films and scan the phosphoimages for these experiments. He said when Xiaoming told him over the phone that these controls were blank with no bands, he used the image of AGO1 input blank to temporarily reserve the space for these controls and planned to replace them with the real images after he returned back to the lab. However, he forgot to replace them or tell Xiaoming about it after the operation, otherwise Xiaoming would have helped him finish the figure making. Hongwei feels very sorry about this, and as a PI now himself, he totally understands how much damage such careless mistakes can cause. It is a good lesson for all of us. Best Regards, Hailing Jin ![file] , 
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 Dear Dr. Aneurus Inconstans, Thank you for pointing out mistakes in our article! I double checked the figure panels, and found that these panels are indeed duplicated and mistakenly positioned. I sincerely apologize to you and all the readers for these terrible mistakes. At the meantime, I was thinking how these horrible mistakes could happen at all. I went through my computer and noticed that these mistakes happened in version 5.3. Then I recalled that at that time I was distracted by multiple things, especially about making therapeutic schedule for my daughteru2019s Mitral Valve Repair surgery, which unavoidably defocused me a lot. I spent a lot of time on road and in hospital, and forgot to replace the wrong figures after I got the right figures. Fortunately, after I searching my computer, I found those original pictures. I sent an illustrative PPT with associated original files to Dr. Hailing Jin. I explained the sequence of events to both Hailing and Xiaoming, to whom I owe a lot. Please refer to the illustrative PPT files and associated original files posted by Hailing. We also checked the similar assays with other papers published from our group and other groups, and confirmed that the Tubulin protein (Hu, Zhao et al. 2019) and U6 RNA (Zhu, Hu et al. 2011) were indeed not associated with AtAGO1 and AtAGO2. Thank you again for pointing out the mistakes that I otherwise would never notice. I sincerely apologized to you, all readers, and all collaborators for these mistakes! Best, Hongwei 1.tHu, P., H. Zhao, P. Zhu, Y. Xiao, W. Miao, Y. Wang and H. Jin (2019).  Dual regulation of Arabidopsis AGO2 by arginine methylation. Nature communications 10(1): 844. 2.tZhu, H., F. Hu, R. Wang, X. Zhou, S. H. Sze, L. W. Liou, A. Barefoot, M. Dickman and X. Zhang (2011).  Arabidopsis Argonaute10 specifically sequesters miR166 165 to regulate shoot apical meristem development. Cell 145(2): 242-256. , 
 #2 Dear Dr. Jin, thank you for you reply. I am sorry to hear about the health issue of Dr. Zhao s daughter, I hope everything is fine by now. As corresponding author of the paper, could you please clarify whether you are going to contact the journal in order to submit a correction as soon as possible? , 
 Sure, we will do that. Hailing , 
 One and a half years later, no correction was issued yet, despite authors promised so. Any comment update? , 
 I will check with the editorial office again. ,