Rotavirus replication is correlated with S/G2 interphase arrest of the host cell cycle

Abstract

In infected cells rotavirus (RV) replicates in viroplasms, cytosolic structures that require a stabilized microtubule (MT) network for their assembly, maintenance of the structure and perinuclear localization. Therefore, we hypothesized that RV could interfere with the MT-breakdown that takes place in mitosis during cell division. Using synchronized RV-permissive cells, we show that RV infection arrests the cell cycle in S/G2 phase, thus favoring replication by improving viroplasms formation, viral protein translation, and viral assembly. The arrest in S/G2 phase is independent of the host or viral strain and relies on active RV replication. RV infection causes cyclin B1 down-regulation, consistent with blocking entry into mitosis. With the aid of chemical inhibitors, the cytoskeleton network was linked to specific signaling pathways of the RV-induced cell cycle arrest. We found that upon RV infection Eg5 kinesin was delocalized from the pericentriolar region to the viroplasms. We used a MA104-Fucci system to identify three RV proteins (NSP3, NSP5, and VP2) involved in cell cycle arrest in the S-phase. Our data indicate that there is a strong correlation between the cell cycle arrest and RV replication.

Conserved Rotavirus NSP5 and VP2 Domains Interact and Affect Viroplasm

“…A clear example is VP2, which is well characterized as the RV core shell encapsidating the viral genome (2,32,33,54,55), acting as a cofactor of RdRp VP1 (28,30,39) and anchoring VP6 to form DLPs (34,55). Aside from its structural functions, VP2 has other roles during the RV life cycle, such as permitting perinuclear motion of viroplasms (10) or being associated with the RV-induced cell cycle arrest (56). Another exciting aspect is its role as the main building block for viroplasm formation.…”

A Host Factor GPNMB Restricts Porcine Circovirus Type 2 (PCV2) Replication and Interacts With PCV2 ORF5 Protein

“…It has been generally accepted that viruses could regulate host cellular life cycle to favor their replication (Tischer et al, 1987). Rotavirus replication correlated with S/G2 interphase arrest in cell cycle (Gluck et al, 2017). Herpesviruses (Flemington, 2001), severe acute respiratory syndrome coronavirus (SARS-CoV) protein (Yuan et al, 2005), influenza A virus and its NS1 protein (Jiang et al, 2013), human respiratory syncytial virus and murine norovirus (MNV) (Davies et al, 2015) can induce cell cycle arrest in the G0/G1 phase.…”

Treading a HOSTile path: Mapping the dynamic landscape of host cell–rotavirus interactions to explore novel host-directed curative dimensions

“…An intersection between RV infection and host cell cycle machinery has been reported from two independent studies. Both the studies concluded arrest of the cell cycle at S phase to produce an environment conducive to rotaviral replication independent of the RV strains and RV-permissive cell lines [ 235 , 238 ]. Prolonged intra-S phase retention observed in the presence of actively replicating RV is enabled by i) heralding the G1-S phase transition [ 235 ] and also by ii) impending entry into the M phase ( Figure 4b ) [ 238 ].…”

“…Both the studies concluded arrest of the cell cycle at S phase to produce an environment conducive to rotaviral replication independent of the RV strains and RV-permissive cell lines [ 235 , 238 ]. Prolonged intra-S phase retention observed in the presence of actively replicating RV is enabled by i) heralding the G1-S phase transition [ 235 ] and also by ii) impending entry into the M phase ( Figure 4b ) [ 238 ]. Pro-viral implications of prolonged S phase duration can be multifaceted such as stabilization of microtubular network for maintenance and maturation of viroplasmic structures, increase in host replication proteins which can potentially be usurped during viral replication, and generation of anti-apoptotic environment to ensure completion of viral replication cycle [ 235 , 238 ].…”