Abstract
Kitaviridae, a newly recognized virus family, includes plant viruses infecting crops of great global importance, notably citrus. Despite its significant impact on citrus agricultural production, the molecular mechanisms underlying kitavirus infections remain largely unknown. Here, we engineered a recombinant citrus leprosis virus C (CiLV-C, genus Cilevirus) expressing green fluorescent protein (GFP) and demonstrated its feasibility for studying the biology of cilevirus. Genetic manipulation of rCiLV-C-GFP revealed that vRNA1 is essential for replication and can self-replicate independently, while vRNA2 is crucial for movement. The intergenic region between the polymerase and capsid protein (CP) acts as a promoter for CP gene expression. Frameshift and deletion analyses provided key insights into replication, movement, and morphogenesis. We reported that CP is critical for viral RNA accumulation, while movement protein (p32) facilitates viral spread. The putative glycoprotein (p61) is not structurally essential, as its deletion did not affect virion assembly, whereas the putative matrix protein (p24) is critical for morphogenesis, likely acting as a structural protein. Deletion of the RNA silencing suppressor (RSS, p15) and p15-p61 attenuated symptoms, implicating them as virulence factors. Additional analyses revealed that CP enhances vRNA accumulation through a mechanism independent of RSS. CP exhibits RNA-binding properties and interacts with eukaryotic initiation factor 4A (eIF4A), suggesting a role in translation. Overexpression of eIF4A increased CiLV-C RNA accumulation, while eIF4A knockdown reduced it, indicating that CP may recruit eIF4A to promote replication. Similar results were observed with turnip crinkle virus (TCV), and notably, the TCV CP efficiently restored RNA accumulation of a CP-defective CiLV-C, suggesting the existence of a conserved, CP-dependent, replication-related mechanism shared across distinct virus families. Our findings support the proposal of an initial model that elucidates the mechanism through which the CPs drive the production of high levels of vRNA manipulating host eIFs.
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Claims 16
Claim · model-written summary
A recombinant GFP-expressing CiLV-C was feasible for cilevirus research.
CitrusOrganisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“we engineered a recombinant citrus leprosis virus C (CiLV-C, genus Cilevirus) expressing green fluorescent protein (GFP) and demonstrated its feasibility for studying the biology of cilevirus.”
Claim · model-written summary
Viral RNA1 was essential for replication and independently self-replicated.
Verbatim
e0“Genetic manipulation of rCiLV-C-GFP revealed that vRNA1 is essential for replication and can self-replicate independently”
t0“plant virus”
Claim · model-written summary
Viral RNA2 was crucial for movement.
Verbatim
e0“while vRNA2 is crucial for movement.”
t0“plant virus”
Claim · model-written summary
The intergenic region promoted CP gene expression.
Verbatim
e0“The intergenic region between the polymerase and capsid protein (CP) acts as a promoter for CP gene expression.”
t0“plant virus”
Claim · model-written summary
CP was critical for viral RNA accumulation.
Verbatim
e0“CP is critical for viral RNA accumulation”
t0“plant virus”
Claim · model-written summary
Movement protein p32 facilitated viral spread.
Verbatim
e0“movement protein (p32) facilitates viral spread.”
t0“plant virus”
Claim · model-written summary
Deleting p61 did not affect virion assembly.
- Taxon
- plant virust0
- Treatment
- its deletione0
- Gene
- putative glycoprotein (p61)e0
- Outcome
- virion assemblye0
- Direction
- did not affecte0
Not reportedComparator
Verbatim
e0“the putative glycoprotein (p61) is not structurally essential, as its deletion did not affect virion assembly”
t0“plant virus”
Claim · model-written summary
The putative matrix protein p24 was critical for virion morphogenesis.
Verbatim
e0“the putative matrix protein (p24) is critical for morphogenesis, likely acting as a structural protein.”
t0“plant virus”
Claim · model-written summary
Deleting p15 attenuated disease symptoms.
- Taxon
- plant virust0
- Treatment
- Deletion of the RNA silencing suppressor (RSS, p15)e0
- Gene
- RSS, p15e0
- Outcome
- symptomse0
- Direction
- attenuatede0
Not reportedComparator
Verbatim
e0“Deletion of the RNA silencing suppressor (RSS, p15) and p15-p61 attenuated symptoms”
t0“plant virus”
Claim · model-written summary
Deleting p15-p61 attenuated symptoms.
Not reportedComparator
Verbatim
e0“Deletion of the RNA silencing suppressor (RSS, p15) and p15-p61 attenuated symptoms”
t0“plant virus”
Claim · model-written summary
CP enhanced viral RNA accumulation independently of RSS.
Verbatim
e0“CP enhances vRNA accumulation through a mechanism independent of RSS.”
t0“plant virus”
Claim · model-written summary
CP bound RNA and interacted with eIF4A.
Verbatim
e0“CP exhibits RNA-binding properties and interacts with eukaryotic initiation factor 4A (eIF4A)”
t0“plant virus”
Claim · model-written summary
Overexpressing eIF4A increased CiLV-C RNA accumulation.
- Taxon
- plant virust0
- Treatment
- Overexpression of eIF4Ae0
- Gene
- eIF4Ae0
- Outcome
- CiLV-C RNA accumulatione0
- Direction
- increasede0
Not reportedComparator
Verbatim
e0“Overexpression of eIF4A increased CiLV-C RNA accumulation”
t0“plant virus”
Claim · model-written summary
Knockdown of eIF4A reduced viral RNA accumulation.
Not reportedComparator
Verbatim
e0“while eIF4A knockdown reduced it”
t0“plant virus”
Claim · model-written summary
Turnip crinkle virus showed similar results for eIF4A.
- Taxon
- turnip crinkle virusTurnip crinkle viruse0
- Outcome
- Similar resultse0
Verbatim
e0“Similar results were observed with turnip crinkle virus (TCV)”
Claim · model-written summary
TCV CP restored RNA accumulation in CP-defective CiLV-C.
Verbatim
e0“the TCV CP efficiently restored RNA accumulation of a CP-defective CiLV-C”
t0“plant virus”