Physical association of Arabidopsis hypersensitive induced reaction proteins (HIRs) with the immune receptor RPS2
Authors: Hans Thordal-Christensen
Fumiaki Katagiri
Le V Nguyen
Xia Wang
Jane Glazebrook
Yiping Qi
Angus S Murphy
Kenichi Tsuda
Jinshan Lin
Journal: Journal of Biological Chemistry (2011)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: Arabidopsis; RPS2; AtHIR proteins; effector-triggered immunity; complex components
Affiliations: Department of Plant Biology, Microbial and Plant Genomics Institute, University of Minnesota, St. Paul, Minnesota 55108, USA.

Abstract:  Arabidopsis RPS2 is a typical nucleotide-binding leucine-rich repeat resistance protein, which indirectly recognizes the bacterial effector protein AvrRpt2 and thereby activates effector-triggered immunity (ETI). Previously, we identified two hypersensitive induced reaction (AtHIR) proteins, AtHIR1 (At1g09840) and AtHIR2 (At3g01290), as potential RPS2 complex components. AtHIR proteins contain the stomatin/prohibitin/flotillin/HflK/C domain (also known as the prohibitin domain or band 7 domain). In this study, we confirmed that AtHIR1 and AtHIR2 form complexes with RPS2 in Arabidopsis and Nicotiana benthamiana using a pulldown assay and fluorescence resonance energy transfer (FRET) analysis. Arabidopsis has four HIR family genes (AtHIR1-4). All AtHIR proteins could form homo- and hetero-oligomers in vivo and were enriched in membrane microdomains of the plasma membrane. The mRNA levels of all except AtHIR4 were significantly induced by microbe-associated molecular patterns, such as the bacterial flagellin fragment flg22. Athir2-1 and Athir3-1 mutants allowed more growth of Pto DC3000 AvrRpt2, but not Pto DC3000, indicating that these mutations reduce RPS2-mediated ETI but do not affect basal resistance to the virulent strain. Overexpression of AtHIR1 and AtHIR2 reduced growth of Pto DC3000. Taken together, the results show that the AtHIR proteins are physically associated with RPS2, are localized in membrane microdomains, and quantitatively contribute to RPS2-mediated ETI. 

Retracted: No

PubPeer comments:
 We read,  **Methods** A double-blinded, randomized, prospective, investigational study of *dutasteride* for the treatment of COVID-19 (NCT04446429). Then  **Results** A total of 130 SARS-CoV-2 positive men were included in the study, 64 subjects in the *dutasteride* arm and 66 subjects in the placebo-controlled group. But the [linked clinical trial NCT04446429] describes a trial of **Proxalutamide**, with 262 participants. To the best of my knowledge, these are different androgen blockers. Here is the second author s summary of the results: https: clinicaltrials.gov ProvidedDocs 29 NCT04446429 Prot_SAP_000.pdf.  ...the 262 subjects who received at least 1 dose of either proxalutamide or placebo (134 subjects in the proxalutamide arm and 128 subjects in the placebo arm). Are the authors conducting so many clinical trials in Brazil that the different patients just blur together? UPDATE: when we look at the History section of NCT04446429 in the Clinical Trials registry, it turns out to have been a moving target, initially registered in June 2020 for dutasteride. Then an arm was added to the trial [in July 2020] , on the suggestion of [the manufacturers of proxalutamide] with the intention of treating another 120 patients with that drug. Finally: between 10 and 29 December, towards the end of recruitment, the researchers realised that the patients they had previously treated with dutasteride had in fact been treated with proxalutamide, and the dutasteride arm was dropped. https: clinicaltrials.gov ct2 history NCT04446429?A=5 B=6 C=Side-by-Side#StudyPageTop , 
 As well as morphing from dutasteride to proxalutamide, the description of NCT04446429 was revised to drop the inclusion criterion of androgenic alopecia (male pattern baldness), which was the key rationale for the study; and to drop all but one of the end-point measures of improvement. Another version of this preprint is archived at ResearchGate: [Dutasteride Reduces Time to Remission in COVID-19: Results From a Randomized Double Blind Placebo Controlled Interventional Trial (The DUTA AndroCoV-Trial)] . That version has the same number of cases, same results, same clinical-trial link to NCT04446429. However, that version s title is a clue. There is in fact a Clinical Trial registry entry for a **different** trial, the [EAT-DUTA AndroCoV Trial] (NCT04729491) with 138 cases in total. That trial finished in 7 October 2020, and was retrospectively registered on 28 January 2021. No results are shown. Perhaps the dutasteride patients from NCT04446429 were all reassigned to the newly-created NCT04729491? If that is the explanation for the 130 cases involved in this study, I am wondering about the criteria used to select 66 placebo-arm subjects from NCT04446429 to make the placebo arm of NCT04729491. , 
 See also https: steamtraen.blogspot.com 2021 10 applying-john-carlisles-table-1.html ,