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Novel asymmetric isopeptide bond steers directional genomic RNA egress from icosahedral virus

ABSTRACT: How essential asymmetries are built into icosahedral virus to facilitate directional genome egress is unknown. Here we capture the structure and dynamics of disassembly intermediates of an icosahedral virus. We reveal localized RNA density poised for release beneath 5-fold axes on one half of the viral particle. The asymmetric distribution of RNA is associated with a unique interchain isopeptide bond (Glu-Lys) that flanks RNA interacting loci on the capsid protein N-terminus. This singular isopeptide bond confers an essential 'loaded-die' feature that steers directional genome egress into target host cells.

INSTRUMENT(S):

ORGANISM(S): Turnip Crinkle Virus

TISSUE(S): Leaf

SUBMITTER: Sean Braet

LAB HEAD: Ganesh S. Anand

PROVIDER: PXD067878 | Pride | 2025-11-06

REPOSITORIES: Pride

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Source:

			Action	DRS
	070417_RR_arunregeneratedcolumn_Native_UnD1_IA_final_peptide.csv	Csv
	070417_RR_arunregeneratedcolumn_Native_UnD2_IA_final_peptide.csv	Csv
	070417_RR_arunregeneratedcolumn_Native_UnD3_IA_final_peptide.csv	Csv
	100417_RR_denvcolumn_TCV_Native_UnD1_IA_final_peptide.csv	Csv
	100417_RR_denvcolumn_TCV_Native_UnD2_IA_final_peptide.csv	Csv

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Project description:Collagen deposition is a key process during idiopathic pulmonary fibrosis (IPF); however, little is known about the dynamics of collagen formation during disease development. Tissue samples of early stages of human disease are not readily available and it is difficult to identify changes in collagen content, since standard collagen analysis does not distinguish between 'old' and 'new' collagen. Therefore, the current study aimed to (i) investigate the dynamics of new collagen formation in mice using bleomycin-induced lung fibrosis in which newly synthesized collagen was labelled with deuterated water and (ii) use this information to identify genes and processes correlated to new collagen formation from gene expression analysis. Lung fibrosis was induced in female C57BL/6 mice by bleomycin instillation and sacrificed. Animals were sacrificed at 1 to 5 weeks after fibrosis induction. Collagen synthesized during the week before sacrifice was labelled with deuterium by providing mice with deuterated drinking water. After sacrifice, lung tissue was collected for microarray analysis, determination of new collagen formation, and histology. Deuterated water labelling showed a strong increase in new collagen formation already during the first week after fibrosis induction and a complete return to baseline at five weeks. Correlation of new collagen formation data with gene expression data revealed fibrosis specific processes, of which proliferation was an unexpected one. This was confirmed by measuring cell proliferation and collagen synthesis simultaneously using deuterated water incorporation. Furthermore, new collagen formation strongly correlated with gene expression of e.g. elastin, tenascin C, MMP-14, lysyl oxidase, and type V collagen. These data demonstrate, using a novel combination of technologies, that proliferation and extracellular matrix production are correlated to the core process of fibrosis, i.e. the formation of new collagen. In addition, it identified genes directly correlated to fibrosis, thus providing more insight into the aetiology of IPF. Total RNA was obtained from mouse lungs at timepoint 0 as a control (n = 7) or timepoints 1 (n = 7), 2 (n = 6), 3 (n = 6), 4 (n = 6) or 5 (n = 6) weeks after bleomycin-instillation to induce lung fibrosis.

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Novel asymmetric isopeptide bond steers directional genomic RNA egress from icosahedral virus

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