Dataset Information

Proteomic analysis of virion and genomic fibre of Mimivirus reunion

ABSTRACT: Mimivirus is the prototype of the Mimiviridae family of giant dsDNA viruses, initially isolated in Acanthamoeba. Little is known about the organization of the viral genome inside the membrane limited nucleoid and whether unpacking or other rearrangements are required prior to early transcription and replication. Here we used Mimivirus reunion (an isolate from La Réunion Island) and showed that in vitro opening of its large icosahedral capsid leads to the release of electron dense, 30 nm diameter rod-shaped objects that appear to be expelled and unwinding from the particles. We developed a purification procedure and performed a mass spectrometry-based characterization of the protein content of the genomic fibre, as well as total virion.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Acanthamoeba Castellanii Mimivirus

SUBMITTER: Yohann Couté

LAB HEAD: Virginie Brun

PROVIDER: PXD021585 | Pride | 2022-07-29

REPOSITORIES: Pride

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Publications

The giant mimivirus 1.2 Mb genome is elegantly organized into a 30-nm diameter helical protein shield.

Villalta Alejandro A Schmitt Alain A Estrozi Leandro F LF Quemin Emmanuelle R J ERJ Alempic Jean-Marie JM Lartigue Audrey A Pražák Vojtěch V Belmudes Lucid L Vasishtan Daven D Colmant Agathe M G AMG Honoré Flora A FA Couté Yohann Y Grünewald Kay K Abergel Chantal C

eLife 20220728

Mimivirus is the prototype of the <i>Mimiviridae</i> family of giant dsDNA viruses. Little is known about the organization of the 1.2 Mb genome inside the membrane-limited nucleoid filling the ~0.5 µm icosahedral capsids. Cryo-electron microscopy, cryo-electron tomography, and proteomics revealed that it is encased into a ~30-nm diameter helical protein shell surprisingly composed of two GMC-type oxidoreductases, which also form the glycosylated fibrils decorating the capsid. The genome is arran ...[more]

PMID: 35900198

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Project description:Comparative analysis of transcriptome profiles of G. arboreum L. cv. and its fuzzy-lintless mutant (ANOI 1960) at 0 and 10 dpa. Cotton is one of the most commercially important fibre crops in the world and used as a source for natural textile fibre and cottonseed oil. The fuzzy-lintless ovules of cotton mutants are ideal source for identifying genes involved in fibre development by comparing with fibre bearing ovules of wild-type. To decipher molecular mechanisms involved in fibre cell development, transcriptome analysis has been carried out by comparing G. arboreum cv. (wild-type) with its fuzzy-lintless mutant (ANOI 1960). Fuzzed-lintless mutant line was generated by back cross breeding between FL and Fl (recurrent parent) lines (personal communication by Dr. I. S. Katageri). Basically Fibre less type was a RIL, first recovered from cross between G.arboreum (linted) and G. anomalum (lint less). This RIL was used as donor parent and crossed with normal arboreum (as recurrent parent) to develop G. arboreum FL and G. arboreum Fl isogenic lines. This G. arboreum Fl line is named as ANOI 1960. Cotton bolls were collected at fibre initiation (0 dpa/days post anthesis) and elongation (10 dpa) and gene expression profiles were analyzed in wild-type and ANOI 1960 mutant using Affymetrix cotton GeneChip Genome array. Cotton plants were grown under field condition. Flowers were tagged and cotton bolls were collected during fibre development stages. Total RNA was isolated from fibre bearing ovules of wild-type (WT) and fuzzy-lintless ovules of mutant (ANOI 1960) collected at various (0 and 10 dpa) fibre development stages using SpectrumTM Plant Total RNA kit (Sigma, USA) according to the manufacturerM-bM-^@M-^Ys protocol. Affymetrix cotton GeneChip Genome array (Affymetrix, USA) having 23,977 probe sets representing 21,854 cotton transcripts was used for transcriptome analysis. Three biological replicates were maintained to test the reproducibility and quality of the chip hybridization. cDNA labeling, array hybridization, staining and washing procedures were carried out as described in the Affymetrix protocols. CEL files having estimated probe intensity values were analyzed with GeneSpring GX-11.5 software (Agilent Technologies, USA) to get differentially expressed transcripts. The Robust Multiarray Average (RMA) algorithm was used for the back ground correction, quantile normalization and median polished probe set summarization to generate single expression value for each probe set. Normalized expression values were log2-transformed and differential expression analysis was performed using unpaired t-test. The p-values were corrected by applying the false discovery rate (FDR) correction (Benjamini and Hochberg, 2000).

			Action	DRS
	Genomic_Fibre_R1.mgf	Mgf
	Genomic_Fibre_R1.mzid.gz	Mzid
	Genomic_Fibre_R1.raw	Raw
	Genomic_Fibre_R2.mgf	Mgf
	Genomic_Fibre_R2.mzid.gz	Mzid