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Regulation of coronaviral poly(A) tail length during infection is not coronavirus species- or host cell-specific.

ABSTRACT: It has been demonstrated that the length of the poly(A) tail in the bovine coronavirus (BCoV), which belongs to genus betacoronaviruses, is regulated throughout infection in human rectal tumor-18 (HRT-18) cells, and the length of the poly(A) tail is associated with the efficiency of virus translation. Here, we examined whether the regulation of viral poly(A) tail length is cell-type independent and whether it is a common feature of coronaviruses to assess the significance of the regulation. By ligating head-to-tail viral RNA positive strands and sequencing, we found that (1) the regulation pattern of coronaviral poly(A) tail length in BCoV-infected hamster kidney-21 (BHK-21) cells was similar to that in BCoV-infected HRT-18 cells and (2) the poly(A) tail length of wild-type avian infectious bronchitis virus (IBV) virulent strain IBV-TW1, which is in the genus gammacoronaviruses, varied throughout infection in primary chicken embryo kidney (CEK) cells and in the tracheas of 1-day-old chicks. Interestingly, the poly(A) tail length variation was similarly found in the avirulent IBV strain H120 in CEK cells, although the overall poly(A) tail length was shorter for this virus. The results suggest that the regulation of coronaviral poly(A) tail length during infection may be a common feature among coronaviruses and can occur in a noncancerous cell line (BHK-21 cells), primary cell culture (CEK cells), and living system (chickens), further reinforcing the biological significance of this regulation during coronavirus infection.

SUBMITTER: Shien JH

PROVIDER: S-EPMC7089208 | biostudies-literature | 2014 Dec

REPOSITORIES: biostudies-literature

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Regulation of coronaviral poly(A) tail length during infection is not coronavirus species- or host cell-specific.

Shien Jui-Hung JH Su Yi-Da YD Wu Hung-Yi HY

Virus genes 20140718 3

It has been demonstrated that the length of the poly(A) tail in the bovine coronavirus (BCoV), which belongs to genus betacoronaviruses, is regulated throughout infection in human rectal tumor-18 (HRT-18) cells, and the length of the poly(A) tail is associated with the efficiency of virus translation. Here, we examined whether the regulation of viral poly(A) tail length is cell-type independent and whether it is a common feature of coronaviruses to assess the significance of the regulation. By l ...[more]

PMID: 25034371

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Project description:In the present study, we investigated the roles of interactions among the poly(A) tail, coronavirus nucleocapsid (N) protein, and poly(A)-binding protein (PABP) in the regulation of coronavirus gene expression. Through dissociation constant (Kd ) comparison, we found that the coronavirus N protein can bind to the poly(A) tail with high affinity, establishing N protein as a PABP. A subsequent analysis with UV cross-linking and immunoprecipitation revealed that the N protein is able to bind to the poly(A) tail in infected cells. Further examination demonstrated that poly(A) tail binding by the N protein negatively regulates translation of coronaviral RNA and host mRNA both in vitro and in cells. Although the N protein can interact with PABP and eukaryotic initiation factor 4G (eIF4G), the poor interaction efficiency between the poly(A)-bound N protein and eIF4E may explain the observed decreased translation efficiency. In addition to interaction with translation factor eIF4G, the N protein is able to interact with coronavirus nonstructural protein 9 (nsp9), a replicase protein required for replication. The study demonstrates interactions among the poly(A) tail, N protein, and PABP both in vitro and in infected cells. Of the interactions, binding of the poly(A) tail to N protein decreases the interaction efficiency between the poly(A) tail and eIF4E, leading to translation inhibition. The poly(A)-dependent translation inhibition by N protein has not been previously demonstrated and thus extends our understanding of coronavirus gene expression.IMPORTANCE Gene expression in coronavirus is a complicated and dynamic process. In this study, we demonstrated that coronavirus N protein is able to bind to the poly(A) tail with high affinity, establishing N protein as a PABP. We also show how the interplay between coronavirus 3' poly(A) tail, PABP, and N protein regulates gene expression of the coronavirus and host cell. Of the interactions, poly(A) tail binding by the N protein negatively regulates translation, and to our knowledge, this inhibition of translation by binding of the N protein to poly(A) tail has not been previously studied. Accordingly, the study provides fundamental molecular details regarding coronavirus infection and expands our knowledge of coronavirus gene expression.

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Regulation of coronaviral poly(A) tail length during infection is not coronavirus species- or host cell-specific.

Publications

Regulation of coronaviral poly(A) tail length during infection is not coronavirus species- or host cell-specific.

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