There is an urgent need for vaccines against coronavirus disease 2019 (COVID-19) because of the ongoing SARS-CoV-2 pandemic. Among all approaches, a messenger RNA (mRNA)-based vaccine has emerged as ...a rapid and versatile platform to quickly respond to this challenge. Here, we developed a lipid nanoparticle-encapsulated mRNA (mRNA-LNP) encoding the receptor binding domain (RBD) of SARS-CoV-2 as a vaccine candidate (called ARCoV). Intramuscular immunization of ARCoV mRNA-LNP elicited robust neutralizing antibodies against SARS-CoV-2 as well as a Th1-biased cellular response in mice and non-human primates. Two doses of ARCoV immunization in mice conferred complete protection against the challenge of a SARS-CoV-2 mouse-adapted strain. Additionally, ARCoV is manufactured as a liquid formulation and can be stored at room temperature for at least 1 week. ARCoV is currently being evaluated in phase 1 clinical trials.
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•Development of LNP-encapsulated mRNA vaccine (ARCoV) targeting the RBD of SARS-CoV-2•ARCoV induces neutralizing antibodies and T cell immunity in mice and NHPs•ARCoV vaccination confers full protection against SARS-CoV-2 challenge in mice•ARCoV is a thermostable vaccine candidate for phase I studies
ARCoV is an LNP-encapsulated mRNA vaccine platform that is highly immunogenic and safe in mice and non-human primates, conferring protection against challenge with a SARS-CoV-2 mouse-adapted strain.
SUMMARY
Drought stress induces anthocyanin biosynthesis in many plant species, but the underlying molecular mechanism remains unclear. Ethylene response factors (ERFs) play key roles in plant growth ...and various stress responses, including affecting anthocyanin biosynthesis. Here, we characterized an ERF protein, MdERF38, which is involved in drought stress‐induced anthocyanin biosynthesis. Biochemical and molecular analyses showed that MdERF38 interacted with MdMYB1, a positive modulator of anthocyanin biosynthesis, and facilitated the binding of MdMYB1 to its target genes. Therefore, MdERF38 promoted anthocyanin biosynthesis in response to drought stress. Furthermore, we found that MdBT2, a negative modulator of anthocyanin biosynthesis, decreased MdERF38‐promoted anthocyanin biosynthesis by accelerating the degradation of the MdERF38 protein. In summary, our data provide a mechanism for drought stress‐induced anthocyanin biosynthesis that involves dynamic modulation of MdERF38 at both transcriptional and post‐translational levels.
Significance Statement
MdERF38 promotes anthocyanin biosynthesis by interacting with MdMYB1 and enhancing the binding of MdMYB1 to its target genes in response to drought stress. MdBT2 decreases drought‐induced anthocyanin accumulation by accelerating the degradation of MdERF38.
Summary
Cold stress severely affects plant growth and yield. C‐repeat binding factors (CBFs) play important roles in the response to cold stress. In the present study, we identified an R2R3‐MYB ...transcription factor (TF) MdMYB23 from apple (Malus × domestic) using transcriptome analyses, which was notably induced in response to cold stress. Transgenic apple calli and Arabidopsis with overexpression of MdMYB23 exhibited increased cold tolerance. Electrophoretic mobility shift assay (EMSA) and transient expression assays indicated that MdMYB23 directly bound to the promoters of MdCBF1 and MdCBF2 and activated their expression. MdMYB23 interacted with the promoter of MdANR, a key modulator of proanthocyanidin biosynthesis, and activated its expression to promote proanthocyanidin accumulation and reactive oxygen species (ROS) scavenging. MdBT2 was identified as an MdMYB23‐interacting protein using yeast two‐hybrid (Y2H), pull‐down, and bimolecular fluorescence complementation (BiFC) assays. MdBT2 repressed cold tolerance and proanthocyanidin accumulation by promoting the degradation of MdMYB23 protein. Our findings shed light on the functions of MYB TFs and underlying mechanism in the modulation of plant cold tolerance.
Significance Statement
An apple R2R3‐MYB TF MdMYB23 increases cold tolerance and proanthocyanidin accumulation by directly activating the expression of MdCBF1/2 and MdANR. MdBT2 represses cold tolerance and proanthocyanidin accumulation by promoting the degradation of MdMYB23 protein.
Summary
The plant hormone jasmonic acid (JA) is involved in the cold stress response, and the inducer of CBF expression 1 (ICE1)‐ C‐repeat binding factor (CBF) regulatory cascade plays a key role in ...the regulation of cold stress tolerance. In this study, we showed that a novel B‐box (BBX) protein MdBBX37 positively regulates JA‐mediated cold‐stress resistance in apple.
We found that MdBBX37 bound to the MdCBF1 and MdCBF4 promoters to activate their transcription, and also interacted with MdICE1 to enhance the transcriptional activity of MdICE1 on MdCBF1, thus promoting its cold tolerance.
Two JA signaling repressors, MdJAZ1 and MdJAZ2 (JAZ, JAZMONATE ZIM‐DOMAIN), interacted with MdBBX37 to repress the transcriptional activity of MdBBX37 on MdCBF1 and MdCBF4, and also interfered with the interaction between MdBBX37 and MdICE1, thus negatively regulating JA‐mediated cold tolerance. E3 ligase MdMIEL1 (MIEL1, MYB30‐Interacting E3 Ligase1) reduced MdBBX37‐improved cold resistance by mediating ubiquitination and degradation of the MdBBX37 protein.
The data reveal that MIEL1 and JAZ proteins co‐regulate JA‐mediated cold stress tolerance through the BBX37‐ICE1‐CBF module in apple. These results will aid further examination of the post‐translational modification of BBX proteins and the regulatory mechanism of JA‐mediated cold stress tolerance.
The molecular mechanism of ABA‐promoted anthocyanin accumulation and fruit coloration is less known. Here, an apple bZIP transcription factor MdbZIP44 was identified to be a positive regulator in ...ABA‐promoted anthocyanin accumulation by interacting with MdMYB1 and enhancing its binding capacity to the promoters of downstream target genes. MdBT2 decreased ABA‐promoted anthocyanin accumulation by degrading MdbZIP44 protein.
Phytohormone abscisic acid (ABA) induces anthocyanin biosynthesis; however, the underlying molecular mechanism is less known. In this study, we found that the apple MYB transcription factor MdMYB1 activated anthocyanin biosynthesis in response to ABA. Using a yeast screening technique, we isolated MdbZIP44, an ABA‐induced bZIP transcription factor in apple, as a co‐partner with MdMYB1. MdbZIP44 promoted anthocyanin accumulation in response to ABA by enhancing the binding of MdMYB1 to the promoters of downstream target genes. Furthermore, we identified MdBT2, a BTB protein, as an MdbZIP44‐interacting protein. A series of molecular, biochemical, and genetic analysis suggested that MdBT2 degraded MdbZIP44 protein through the Ubiquitin‐26S proteasome system, thus inhibiting MdbZIP44‐modulated anthocyanin biosynthesis. Taken together, we reveal a novel working mechanism of MdbZIP44‐mediated anthocyanin biosynthesis in response to ABA.
Wounding stress leads to anthocyanin accumulation. However, the underlying molecular mechanism remains elusive. In this study, MdWRKY40 was found to promote wounding-induced anthocyanin biosynthesis ...in association with MdMYB1 and undergo MdBT2-mediated degradation in apple.
We found that MdMYB1, a positive regulator of anthocyanin biosynthesis, was essential for the wounding-induced anthocyanin biosynthesis in apple. MdWRKY40 was identified as an MdMYB1-interacting protein, and enhanced the binding of MdMYB1 to its target genes in response to wounding.
We found that MdBT2 interacted physically with MdWRKY40 and was involved in its degradation through the 26S proteasome pathway.
Our results demonstrate that MdWRKY40 is a key modulator in the wounding-induced anthocyanin biosynthesis, which provides new insights into the regulation of wounding-induced anthocyanin biosynthesis at both the transcriptional and post-translational levels in apple.
Since December 2019, a novel coronavirus SARS-CoV-2 has emerged and rapidly spread throughout the world, resulting in a global public health emergency. The lack of vaccine and antivirals has brought ...an urgent need for an animal model. Human angiotensin-converting enzyme II (ACE2) has been identified as a functional receptor for SARS-CoV-2. In this study, we generated a mouse model expressing human ACE2 (hACE2) by using CRISPR/Cas9 knockin technology. In comparison with wild-type C57BL/6 mice, both young and aged hACE2 mice sustained high viral loads in lung, trachea, and brain upon intranasal infection. Although fatalities were not observed, interstitial pneumonia and elevated cytokines were seen in SARS-CoV-2 infected-aged hACE2 mice. Interestingly, intragastric inoculation of SARS-CoV-2 was seen to cause productive infection and lead to pulmonary pathological changes in hACE2 mice. Overall, this animal model described here provides a useful tool for studying SARS-CoV-2 transmission and pathogenesis and evaluating COVID-19 vaccines and therapeutics.
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•Human ACE2 knockin mice were generated by using CRISPR/Cas9 technology•SARS-CoV-2 leads to robust replication in lung, trachea, and brain•SARS-CoV-2 causes interstitial pneumonia and elevated cytokine in aged hACE2 mice•High dose of SARS-CoV-2 can establish infection via intragastric route in hACE2 mice
The COVID-19 pandemic has brought an urgent need for small animal models. Here, Sun et al. established an ACE2 humanized mouse by CRISPR/Cas9 knockin technology. These hACE2 mice are susceptible to SARS-CoV-2 infection upon intranasal inoculation, and the resulting pulmonary infection and pathological changes resemble those observed in COVID-19 patients.
Summary
MYB transcription factors (TFs) have been demonstrated to play diverse roles in plant growth and development through interaction with basic helix‐loop‐helix (bHLH) TFs. MdbHLH33, an apple ...bHLH TF, has been identified as a positive regulator in cold tolerance and anthocyanin accumulation by activating the expressions of MdCBF2 and MdDFR. In the present study, a MYB TF MdMYB308L was found to also positively regulate cold tolerance and anthocyanin accumulation in apple. We found that MdMYB308L interacted with MdbHLH33 and enhanced its binding to the promoters of MdCBF2 and MdDFR. In addition, an apple RING E3 ubiquitin ligase MYB30‐INTERACTING E3 LIGASE 1 (MdMIEL1) was identified to be an MdMYB308L‐interacting protein and promoted the ubiquitination degradation of MdMYB308L, thus negatively regulated cold tolerance and anthocyanin accumulation in apple. These results suggest that MdMYB308L acts as a positive regulator in cold tolerance and anthocyanin accumulation in apple by interacting with MdbHLH33 and undergoes MdMIEL1‐mediated protein degradation. The dynamic change in MYB‐bHLH protein complex seems to play a key role in the regulation of plant growth and development.
To discover new drugs to combat COVID-19, an understanding of the molecular basis of SARS-CoV-2 infection is urgently needed. Here, for the first time, we report the crucial role of cathepsin L ...(CTSL) in patients with COVID-19. The circulating level of CTSL was elevated after SARS-CoV-2 infection and was positively correlated with disease course and severity. Correspondingly, SARS-CoV-2 pseudovirus infection increased CTSL expression in human cells in vitro and human ACE2 transgenic mice in vivo, while CTSL overexpression, in turn, enhanced pseudovirus infection in human cells. CTSL functionally cleaved the SARS-CoV-2 spike protein and enhanced virus entry, as evidenced by CTSL overexpression and knockdown in vitro and application of CTSL inhibitor drugs in vivo. Furthermore, amantadine, a licensed anti-influenza drug, significantly inhibited CTSL activity after SARS-CoV-2 pseudovirus infection and prevented infection both in vitro and in vivo. Therefore, CTSL is a promising target for new anti-COVID-19 drug development.
SUMMARY
Jasmonic acid (JA) induces chlorophyll degradation and leaf senescence. B‐box (BBX) proteins play important roles in the modulation of leaf senescence, but the molecular mechanism of BBX ...protein‐mediated leaf senescence remains to be further studied. Here, we identified the BBX protein MdBBX37 as a positive regulator of JA‐induced leaf senescence in Malus domestica (apple). Further studies showed that MdBBX37 interacted with the senescence regulatory protein MdbHLH93 to enhance its transcriptional activation on the senescence‐associated gene MdSAG18, thereby promoting leaf senescence. Moreover, the JA signaling repressor MdJAZ2 interacted with MdBBX37 and interfered with the interaction between MdBBX37 and MdbHLH93, thereby negatively mediating MdBBX37‐promoted leaf senescence. In addition, the E3 ubiquitin ligase MdSINA3 delayed MdBBX37‐promoted leaf senescence through targeting MdBBX37 for degradation. The MdJAZ2‐MdBBX37‐MdbHLH93‐MdSAG18 and MdSINA3‐MdBBX37 modules realized the precise modulation of JA on leaf senescence. In parallel, our data demonstrate that MdBBX37 was involved in abscisic acid (ABA)‐ and ethylene‐mediated leaf senescence through interacting with the ABA signaling regulatory protein MdABI5 and ethylene signaling regulatory protein MdEIL1, respectively. Taken together, our results not only reveal the role of MdBBX37 as an integration node in JA‐, ABA‐ and ethylene‐mediated leaf senescence, but also provide new insights into the post‐translational modification of BBX proteins.
Significance Statement
BBX37 interacted with the senescence regulatory protein bHLH93 to enhance its transcriptional activation on the senescence‐associated gene SAG18, thereby promoting leaf senescence. JAZ2 interacted with BBX37 and interfered with the interaction between BBX37 and bHLH93, thereby negatively mediating BBX37‐promoted leaf senescence. SINA3 delayed the BBX37‐promoted leaf senescence through targeting BBX37 for ubiquitination and degradation.
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