Abstract
Rift Valley fever virus (RVFV) is a negative-sense, tripartite RNA virus that is endemic to Africa and the Arabian Peninsula. It can cause severe disease and mortality in humans and domestic livestock and is a concern for its potential to spread more globally. RVFV’s nucleocapsid protein (N) is an RNA-binding protein that is necessary for viral transcription, replication, and the production of nascent viral particles. We have conducted crosslinking, immunoprecipitation, and sequencing (CLIP-seq) to characterize N interactions with host and viral RNAs during infection. In parallel, to precisely measure intracellular N levels, we employed multiple reaction monitoring mass spectrometry (MRM-MS). Our results show that N binds mostly to host RNAs at early stages of infection, yielding nascent virus particles of reduced infectivity. The expression of N plateaus 10 h post-infection, whereas the intracellular viral RNA concentration continues to increase. Moreover, the virions produced later in infection have higher infectivity. Taken together, the detailed examination of these N–RNA interactions provides insight into how the regulated expression of N and viral RNA produces both infectious and incomplete, noninfectious particles.
Highlights
Rift Valley fever virus (RVFV) is a single-stranded RNA virus that belongs to the orderBunyavirales
We used quantitative assessment of N protein and viral RNA levels along with a complete cataloging of N–RNA interactions to address knowledge gaps in the molecular virology of RVFV infection. These results elucidate the extensive interaction network of N with host and viral RNAs, and the dynamics of N and vRNA levels as a predictor of nascent viral infectivity, and suggest that the formation of incomplete virus particles and collective infection of cells are inherent in the overall virus spread strategy of RVFV
We conducted CLIP-seq to characterize the interactions between N and RNAs during
Summary
Rift Valley fever virus (RVFV) is a single-stranded RNA virus that belongs to the orderBunyavirales. Rift Valley fever virus (RVFV) is a single-stranded RNA virus that belongs to the order. RVFV infects livestock, causing hepatitis, hemorrhage, abortion, and death [2]. The high abortion rate caused by RVFV and resultant loss of livestock represents a significant threat to local economies [3]. Transmission to humans can occur both through mosquito bites and by contact with infected agricultural animals or animal products [4]. Symptoms of RVFV infection in humans vary from mild flu-like illness to vision loss, liver damage, hemorrhagic fever, and death. Recent studies have shown that RVFV infection can be abortive in mice [5], and likely in humans [6,7]. No approved vaccine or treatment is currently available for RVFV infection in humans
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