Comparative in vivo analysis of the nsp15 endoribonuclease of murine, porcine and severe acute respiratory syndrome coronaviruses.
Comparative in vivo analysis of the nsp15 endoribonuclease of murine, porcine and severe acute respiratory syndrome coronaviruses.
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DOI：
10.1016/j.virusres.2012.05.006
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发表时间：
2012-08
期刊：
影响因子：
5
通讯作者：
Zhang, Xuming
中科院分区：
文献类型：
--
作者：
Cao, Jianzhong;Zhang, Xuming
关键词：
► MHV nsp15 is distributed throughout the cytoplasm of virus-infected cells. ► MHV but not SARS-CoV or TGEV nsp15 formed speckles when ectopically expressed in cell. ► Two domains (I and III) within nsp15 are responsible for the speckle formation. ► A novel protein-interaction domain is identified in the C-terminus of MHV nsp15. ► nsp15s from the 3 coronaviruses have differential effects on MHV replication. The purpose of this study was to compare the biochemical and biological properties of nonstructural protein (nsp) 15 among mouse hepatitis virus (MHV), severe acute respiratory syndrome coronavirus (SARS-CoV) and transmissible gastroenteritis virus (TGEV) in virus-infected and ectopically expressed cells. In virus-infected cells, MHV nsp15 distributed unevenly throughout the cytoplasm but predominantly in the perinuclear region. When expressed as N-terminal enhanced green fluorescence protein (EGFP) fusion, it predominantly formed speckles in the cytoplasm. In contrast, SARS-CoV and TGEV EGFP-nsp15s distributed smoothly in the whole cell and did not form speckles. Deletion mapping experiments identified two domains responsible for the speckle formation in MHV EGFP-nsp15: Domain I (aa101–150) and Domain III (aa301–374). Interestingly, Domain II (aa151–250) had an inhibitory effect on Domain III- but not on Domain I-mediated speckle formation. Expression of a small (35aa) sequence in Domain III alone was sufficient to form speckles for all 3 viral nsp15s. However, addition of surrounding sequences in Domain III abolished the speckle formation for TGEV nsp15 but not for MHV and SARS-CoV nsp15s. Further domain swapping experiments uncovered additional speckle-inducing and -suppressive elements in nsp15s of SARS-CoV and TGEV. Homotypic interaction involving Domain III of MHV nsp15 was further demonstrated biochemically. Moreover, the biological functions of the expressed nsp15s were assessed in MHV-infected cells. It was found that the effects of EGFP-nsp15s on MHV replication were both virus species- and nsp15 domain-dependent. Collectively these results thus underscore the differential biochemical and biological functions among the nsp15s of MHV, TGEV and SARS-CoV in host cells.
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影响因子：
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作者：
Ziebuhr J
通讯作者：
Ziebuhr J
影响因子：
5.4
作者：
Shi, ST;Schiller, JJ;Lai, MMC
通讯作者：
Lai, MMC
影响因子：
5.4
作者：
Gosert, R;Kanjanahaluethai, A;Baker, SC
通讯作者：
Baker, SC
影响因子：
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作者：
Harcourt, BH;Jukneliene, D;Baker, SC
通讯作者：
Baker, SC
影响因子：
14.9
作者：
BREDENBEEK, PJ;PACHUK, CJ;SPAAN, WJM
通讯作者：
SPAAN, WJM