TY - JOUR
T1 - Serotype-specific regulation of Dengue Virus NS5 protein subcellular localization
AU - Cheng, Colin Xinru
AU - Tan, Min Jie Alvin
AU - Chan, Kitti Wing Ki
AU - Choy, Milly Ming Ju
AU - Roman, Noelia
AU - Arnold, Daniel D.R.
AU - Bifani, Amanda Makha
AU - Kong, Sean Yao Zu
AU - Bist, Pradeep
AU - Nath, Babu K.
AU - Swarbrick, Crystall M.D.
AU - Forwood, Jade K.
AU - Vasudevan, Subhash G.
N1 - Publisher Copyright:
© 2024 The Authors. Published by American Chemical Society.
PY - 2024/6/14
Y1 - 2024/6/14
N2 - Dengue virus (DENV) nonstructural protein 5 (NS5), consisting of methyltransferase and RNA-dependent RNA polymerase (RdRp) domains, is critical for viral RNA synthesis within endoplasmic reticulum-derived replication complexes in the cytoplasm. However, a significant proportion of NS5 is localized to the nucleus of infected cells for DENV2, 3, and 4, whereas DENV1 NS5 is localized diffusely in the cytoplasm. We still have an incomplete understanding of how the DENV NS5 subcellular localization is regulated. Within NS5, two putative nuclear localization signal (NLS) sequences have been identified: NLSCentral residing in the palm of the RdRp domain as well as the recently discovered NLSC-term residing in the flexible region at the C-terminal of the RdRp domain. We have previously shown that DENV2 NS5 nuclear localization can be significantly reduced by single-point mutations to the NLSC-term. Here, we present biochemical, virological, and structural data demonstrating that the relative importance of either NLS in NS5 nuclear localization is unique to each of the four DENV serotypes. DENV1 NS5′s cytoplasmic localization appears to be due to a functionally weak interaction between its NLSCentral and importin-α (IMPα), while DENV2 NS5 is almost exclusively nuclear through its NLSC-term’s strong interaction with IMPα. Both NLSs of DENV3 NS5 appear to contribute to directing its nuclear localization.
AB - Dengue virus (DENV) nonstructural protein 5 (NS5), consisting of methyltransferase and RNA-dependent RNA polymerase (RdRp) domains, is critical for viral RNA synthesis within endoplasmic reticulum-derived replication complexes in the cytoplasm. However, a significant proportion of NS5 is localized to the nucleus of infected cells for DENV2, 3, and 4, whereas DENV1 NS5 is localized diffusely in the cytoplasm. We still have an incomplete understanding of how the DENV NS5 subcellular localization is regulated. Within NS5, two putative nuclear localization signal (NLS) sequences have been identified: NLSCentral residing in the palm of the RdRp domain as well as the recently discovered NLSC-term residing in the flexible region at the C-terminal of the RdRp domain. We have previously shown that DENV2 NS5 nuclear localization can be significantly reduced by single-point mutations to the NLSC-term. Here, we present biochemical, virological, and structural data demonstrating that the relative importance of either NLS in NS5 nuclear localization is unique to each of the four DENV serotypes. DENV1 NS5′s cytoplasmic localization appears to be due to a functionally weak interaction between its NLSCentral and importin-α (IMPα), while DENV2 NS5 is almost exclusively nuclear through its NLSC-term’s strong interaction with IMPα. Both NLSs of DENV3 NS5 appear to contribute to directing its nuclear localization.
KW - dengue virus serotypes
KW - importin
KW - importin-α cocrystal structure
KW - nuclear localization signal
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U2 - 10.1021/acsinfecdis.4c00054
DO - 10.1021/acsinfecdis.4c00054
M3 - Article
C2 - 38811007
AN - SCOPUS:85195065543
SN - 2373-8227
VL - 10
SP - 2047
EP - 2062
JO - ACS Infectious Diseases
JF - ACS Infectious Diseases
IS - 6
ER -