RESUMEN
Human astrovirus (HAstV) is a global cause of gastroenteritis in infants, the elderly, and the immunocompromised. However, the molecular mechanisms that control its susceptibility are not fully understood, as the functional receptor used by the virus has yet to be identified. Here, a genome-wide CRISPR-Cas9 library screen in Caco2 cells revealed that the neonatal Fc receptor (FcRn) can function as a receptor for classical HAstV (Mamastrovirus genotype 1). Deletion of FCGRT or B2M, which encode subunits of FcRn, rendered Caco2 cells and intestinal organoid cells resistant to HAstV infection. We also showed that human FcRn expression renders non-susceptible cells permissive to viral infection and that FcRn binds directly to the HAstV spike protein. Therefore, our findings provide insight into the entry mechanism of HAstV into susceptible cells. We anticipate that this information can be used to develop new therapies targeting human astroviruses, providing new strategies to treat this global health issue.
RESUMEN
Analysis of 5'-RACE clones revealed two Cyp19 transcript variants (gonad p450 aroms 1 and 2) in the gonads and one Cyp transcript (brain p450 arom) in the brain that differed in their 5'-untranslated region (UTR). Since all cDNAs contained an identical open reading frame, it is considered that while Xenopus aromatase may be encoded by a single Cyp19 gene, it is transcribed by tissue-specific promoters, each of which may be regulated by a distinct set of transcriptional factors. While gonad p450 aroms 1 and 2 and brain p450 arom were expressed in the gonads, brain p450 arom was the predominantly expressed aromatase gene in the brain, with a low expression level of gonad p450 arom 1.
Asunto(s)
Aromatasa/genética , Encéfalo/enzimología , Expresión Génica , Gónadas/enzimología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Cartilla de ADN , ADN Complementario , Datos de Secuencia Molecular , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , XenopusRESUMEN
Investigation by RQ-RT-PCR revealed that transcription of the p450 aromatase gene is activated at stage 50, when sex determination of the female begins, and that aromatase gene expression is also activated by exogenously administrated estradiol. In order to determine the molecular basis underlying the specific activation of aromatase gene expression during sex differentiation and in response to exogenous estradiol, we isolated the 5'-flanking fragment of the gene and characterized the promoter sequence. We demonstrated binding sequences to a specific trans-activating factor upstream of the p450 aromatase promoter II, the cAMP response element binding protein/activating transcription factor family, and steroidogenic factor-1. An estrogen response element half-site sequence that recognize an estrogen receptors, was also found.