m6A mRNA Methylation Regulates Epithelial Innate Antimicrobial Defense Against Cryptosporidial Infection

Xia, Zijie; Xu, Jihao; Lu, Eugene Y. C.; He, Wei; Deng, Silu; Gong, Ai Yu; Strass-Soukup, Juliane; Martins, Gislâine A.; Lu, Guoqing; Chen, Xian Ming

doi:10.3389/fimmu.2021.705232

Cited by 9 publications

(9 citation statements)

References 74 publications

(188 reference statements)

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“…Models of intestinal cryptosporidiosis using intestinal epithelial cell lines and 2D epithelial monolayers were employed as previously described 16,57,58 At various time points after Cryptosporidium or vehicle/drug administration, ileum intestinal tissues (4 cm of small intestine tissue from the ileocecal junction) were collected from the animals (minimum of 6 mice from each group). Biochemical analysis, RT-qPCR, immunofluorescent staining, and histology were performed as previously reported 57,58 . For in vitro and ex vivo infection, cell cultures or 2D monolayers were exposed to C. parvum infection for 4 h and culture medium was changed to remove free parasite.…”

Section: Generation Of Conditional Intestinal Epithelialmentioning

confidence: 99%

“…For quantitative analysis of RNA expression, comparative real-time PCR was performed as previously reported 57,58 monolayers. 2D monolayers were exposed to C. parvum infection for 24 h, in the presence or absence of anti-viral and anti-parasitic drugs or their combination, followed by RT-qPCR measurement of cpHsp70.…”

Section: Rt-qpcrmentioning

confidence: 99%

“…Western blot and dot blot were performed as reported in our previous study 57,58 . Briefly, cell lysates were collected and SDS-PAGE gel electrophoresis was used to separate denatured proteins on 10% TGS gels.…”

Section: Cytoplasmic Extracts Western Blot and Dot Blotmentioning

confidence: 99%

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Cryptosporidium uses CSpV1 to activate host type I interferon and attenuate antiparasitic defenses

Deng

Gong

et al. 2023

Nat Commun

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Cryptosporidium infects gastrointestinal epithelium and is a leading cause of infectious diarrhea and diarrheal-related death in children worldwide. There are no vaccines and no fully effective therapy available for the infection. Type II and III interferon (IFN) responses are important determinants of susceptibility to infection but the role for type I IFN response remains obscure. Cryptosporidium parvum virus 1 (CSpV1) is a double-stranded RNA (dsRNA) virus harbored by Cryptosporidium spp. Here we show that intestinal epithelial conditional Ifnar1−/− mice (deficient in type I IFN receptor) are resistant to C. parvum infection. CSpV1-dsRNAs are delivered into host cells and trigger type I IFN response in infected cells. Whereas C. parvum infection attenuates epithelial response to IFN-γ, loss of type I IFN signaling or inhibition of CSpV1-dsRNA delivery can restore IFN-γ-mediated protective response. Our findings demonstrate that type I IFN signaling in intestinal epithelial cells is detrimental to intestinal anti-C. parvum defense and Cryptosporidium uses CSpV1 to activate type I IFN signaling to evade epithelial antiparasitic response.

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Section: Generation Of Conditional Intestinal Epithelialmentioning

confidence: 99%

Section: Rt-qpcrmentioning

confidence: 99%

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Cryptosporidium uses CSpV1 to activate host type I interferon and attenuate antiparasitic defenses

Deng

Gong

et al. 2023

Nat Commun

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“… 2 , 23 In epithelial cells, ALKBH5 is decreased in response to infection by Streptococcus suis and Cryptosporidium parvum , with the involvement of TLR/MYD88/NF-кB signaling activation. 50 Future study is needed to reveal the mechanisms underlying downregulation of ALKBH5 during bacterial infection, for example, to clarify whether it is due to the activation of TLRs signaling or even reprogramming metabolic status, which will contribute to develop corresponding intervention strategies for bacterial infections.…”

Section: Discussionmentioning

confidence: 99%

m6A demethylase ALKBH5 is required for antibacterial innate defense by intrinsic motivation of neutrophil migration

Liu

Song

Zhao

et al. 2022

Sig Transduct Target Ther

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Neutrophil migration into the site of infection is necessary for antibacterial innate defense, whereas impaired neutrophil migration may result in excessive inflammation and even sepsis. The neutrophil migration directed by extracellular signals such as chemokines has been extensively studied, yet the intrinsic mechanism for determining neutrophil ability to migrate needs further investigation. N6-methyladenosine (m6A) RNA modification is important in immunity and inflammation, and our preliminary data indicate downregulation of RNA m6A demethylase alkB homolog 5 (ALKBH5) in neutrophils during bacterial infection. Whether m6A modification and ALKBH5 might intrinsically modulate neutrophil innate response remain unknown. Here we report that ALKBH5 is required for antibacterial innate defense by enhancing intrinsic ability of neutrophil migration. We found that deficiency of ALKBH5 increased mortality of mice with polymicrobial sepsis induced by cecal ligation and puncture (CLP), and Alkbh5-deficient CLP mice exhibited higher bacterial burden and massive proinflammatory cytokine production in the peritoneal cavity and blood because of less neutrophil migration. Alkbh5-deficient neutrophils had lower CXCR2 expression, thus exhibiting impaired migration toward chemokine CXCL2. Mechanistically, ALKBH5-mediated m6A demethylation empowered neutrophils with high migration capability through altering the RNA decay, consequently regulating protein expression of its targets, neutrophil migration-related molecules, including increased expression of neutrophil migration-promoting CXCR2 and NLRP12, but decreased expression of neutrophil migration-suppressive PTGER4, TNC, and WNK1. Our findings reveal a previously unknown role of ALKBH5 in imprinting migration-promoting transcriptome signatures in neutrophils and intrinsically promoting neutrophil migration for antibacterial defense, highlighting the potential application of targeting neutrophil m6A modification in controlling bacterial infections.

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“…T. brucei has also been shown to use m 6 A marks in poly(A) tails to protect against deadenylation and degradation (Viegas et al, 2022). For parasitic infections involving Cryptosporidium parvum , there is a 30% global increase in the m 6 A methylome of host cells eight hours postinfection due to down regulation of ALKBH5, FTO, and a nuclear factor kappa‐light‐chain‐enhancer of activated B signaling pathway for innate antimicrobial immune defense (Z. Xia, Xu, et al, 2021). Important future areas of study include how widespread METTL3 orthologs are in human parasites and the prevalence of m 6 A methylome changes for host and parasitic RNAs.…”

Section: Roles Of M6a Mtases In Infectious Agents: Viruses and Parasitesmentioning

confidence: 99%

Ghost authors revealed: The structure and function of human N⁶‐methyladenosine RNA methyltransferases

Breger,

Kunkler,

O'Leary

et al. 2023

WIREs RNA

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Despite the discovery of modified nucleic acids nearly 75 years ago, their biological functions are still being elucidated. N6‐methyladenosine (m6A) is the most abundant modification in eukaryotic messenger RNA (mRNA) and has also been detected in non‐coding RNAs, including long non‐coding RNA, ribosomal RNA, and small nuclear RNA. In general, m6A marks can alter RNA secondary structure and initiate unique RNA–protein interactions that can alter splicing, mRNA turnover, and translation, just to name a few. Although m6A marks in human RNAs have been known to exist since 1974, the structures and functions of methyltransferases responsible for writing m6A marks have been established only recently. Thus far, there are four confirmed human methyltransferases that catalyze the transfer of a methyl group from S‐adenosylmethionine (SAM) to the N6 position of adenosine, producing m6A: methyltransferase‐like protein (METTL) 3/METTL14 complex, METTL16, METTL5, and zinc‐finger CCHC‐domain‐containing protein 4. Though the methyltransferases have unique RNA targets, all human m6A RNA methyltransferases contain a Rossmann fold with a conserved SAM‐binding pocket, suggesting that they utilize a similar catalytic mechanism for methyl transfer. For each of the human m6A RNA methyltransferases, we present the biological functions and links to human disease, RNA targets, catalytic and kinetic mechanisms, and macromolecular structures. We also discuss m6A marks in human viruses and parasites, assigning m6A marks in the transcriptome to specific methyltransferases, small molecules targeting m6A methyltransferases, and the enzymes responsible for hypermodified m6A marks and their biological functions in humans. Understanding m6A methyltransferases is a critical steppingstone toward establishing the m6A epitranscriptome and more broadly the RNome.This article is categorized under: RNA Interactions with Proteins and Other Molecules > Protein‐RNA Recognition RNA Interactions with Proteins and Other Molecules > RNA‐Protein Complexes RNA Interactions with Proteins and Other Molecules > Protein‐RNA Interactions: Functional Implications

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m6A mRNA Methylation Regulates Epithelial Innate Antimicrobial Defense Against Cryptosporidial Infection

Cited by 9 publications

References 74 publications

Cryptosporidium uses CSpV1 to activate host type I interferon and attenuate antiparasitic defenses

Cryptosporidium uses CSpV1 to activate host type I interferon and attenuate antiparasitic defenses

m6A demethylase ALKBH5 is required for antibacterial innate defense by intrinsic motivation of neutrophil migration

Ghost authors revealed: The structure and function of human N⁶‐methyladenosine RNA methyltransferases

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m6A mRNA Methylation Regulates Epithelial Innate Antimicrobial Defense Against Cryptosporidial Infection

Cited by 9 publications

References 74 publications

Cryptosporidium uses CSpV1 to activate host type I interferon and attenuate antiparasitic defenses

Cryptosporidium uses CSpV1 to activate host type I interferon and attenuate antiparasitic defenses

m6A demethylase ALKBH5 is required for antibacterial innate defense by intrinsic motivation of neutrophil migration

Ghost authors revealed: The structure and function of human N6‐methyladenosine RNA methyltransferases

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Ghost authors revealed: The structure and function of human N⁶‐methyladenosine RNA methyltransferases