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Differential expression of proteins in hematopoietic stem/progenitor cells lacking the Cullin-5 protein

ABSTRACT: Cullin proteins are scaffolds that coordinate assembly of cullin-RING E3 ubiquitin (Ub) ligases (CRL), complexes that control post-translational ubiquitin modification and degradation of cellular proteins. Cullin-5 (Cul5) coordinates assembly of CRL complexes containing the RING E3 ligase Rbx1/2, the adapter proteins Elongins B and C, and a Suppressor of Cytokine Signalling (SOCS) box-containing substrate recognition protein. To explore potential roles for Cul5, we generated mice lacking Cul5 in the in hematopoietic system. Analyses included biological and molecular studies including proteomic analysis of differential expression of proteins in primary purified hematopoietic stem/progenitor (LSK) cells lacking Cul5.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Stem Cell

SUBMITTER: Laura Dagley

LAB HEAD: Prof. Warren Alexander

PROVIDER: PXD051430 | Pride | 2025-01-26

REPOSITORIES: pride

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Source:

			Action	DRS
	DIANN_libfree_290921.zip	Other
	P4049_10.2_1_1_6085.d.zip	Other
	P4049_11_1_1_6093.d.zip	Other
	P4049_12.2_1_1_6084.d.zip	Other
	P4049_13_1_1_6091.d.zip	Other

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Project description:Oral transmission of T. cruzi is probably the most frequent mechanism among animals in the wild. In this context, there is a high prevalence of human infections in regions where triatomine infection with T. cruzi is low. This led to the hypothesis that the consumption of raw or undercooked meat from animals infected with T. cruzi could be responsible for the transmission of the infection. Therefore, the general objective of this study was to demonstrate the role of meat consumption from infected animals in the oral transmission of T. cruzi infection. Groups of five female mice Balb/c were fed with muscles obtained from mice in the acute phase of infection by the clone H510 C8C3hvir of T. cruzi, and the infection of the fed mice was monitored by a parasitemia curve. Similarly, we assessed the infective capacity of T. cruzi trypomastigotes and amastigotes by infecting groups of five mice Balb/c females, which were infected orally using a nasogastric probe, and the infection was monitored by parasitemia curve. Finally, different trypomastigote and amastigote inoculums were used to determine their infective capacity. Adhesion assays of T. cruzi proteins to AGS stomach cells were performed and the adhered proteins were detected by western blotting using monoclonal or polyclonal antibodies. In all cases, 60–100% of animals were fed meat from mice infected in the acute phase or infected by means of a nasogastric probe with trypomastigotes or amastigotes. These animals developed high parasitemia, and 80% died around day 40 post-infection. The adhesion tests showed that cruzipain is a molecule of trypomastigotes and amastigotes that binds to AGS cells. LC-MS/MS also confirmed that transialidase may be involved in TCT attachment of TCT or invasion of stomach cells. It was concluded that the consumption of meat from infected animals in the acute phase of T. cruzi infection allows transmission of the infection. Similarly, trypomastigotes and amastigotes were able to infect mice when administered orally, while cruzipain was a relevant molecule in this infective process.

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Differential expression of proteins in hematopoietic stem/progenitor cells lacking the Cullin-5 protein

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