Dataset Information

Deep proteome analysis of Leishmania major Friedlin

ABSTRACT: Deep proteome analysis to assess the impact of FUT1 and LPG on the L. major Friedlin proteome

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Leishmania Major Strain Friedlin

SUBMITTER: Nichollas Scott

LAB HEAD: Stephen M. Beverley

PROVIDER: PXD015966 | Pride | 2022-10-12

REPOSITORIES: Pride

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Source:

			Action	DRS
	MQ_output.zip	Other
	Nsco_20180505_LM_FUT_dLFQ_B1_F01.raw	Raw
	Nsco_20180505_LM_FUT_dLFQ_B1_F02.raw	Raw
	Nsco_20180505_LM_FUT_dLFQ_B1_F03.raw	Raw
	Nsco_20180505_LM_FUT_dLFQ_B1_F04.raw	Raw

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Project description:Protein-coding genes in kinetoplastid protists are transcribed from polycistronic arrays, yielding RNA precursors which are processed to form mature transcripts bearing a 5M-bM-^@M-^Y spliced leader (SL) and 3M-bM-^@M-^Y poly(A) tract. Regions of transcription initiation and termination lack known eukaryotic promoter and terminator elements, and current data suggest that transcription is instead regulated predominantly through epigenetic mechanisms. Several epigenetic marks, including histone modifications, histone variants, and an atypical DNA modification known as base J have been localized to regions of transcription initiation or termination in Trypanosoma brucei, Trypanosoma cruzi, and/or Leishmania major. Despite this conservation, the phenotypes of base J mutants vary significantly across trypanosomatids, suggesting that the specific epigenetic networks governing transcription initiation and termination have diverged significantly during evolution. In this light, we sought to characterize and compare the roles of the histone variants H2A.Z, H2B.V, and H3.V in L. major. As in T. brucei, the histone variants H2A.Z and H2B.V were shown to be essential in L. major using a powerful quantitative plasmid segregation-based test. In contrast and again similar to T. brucei, H3.V is not essential in Leishmania as H3.V-null lines grew normally, resembled WT, and remained infectious. Using SL-primed RNA-seq, we found that H3.V-null parasites have steady-state transcript levels comparable to WT parasites and display no defects in the efficiency of transcription termination at convergent strand switch regions (SSRs). Our results show a conservation of histone variant phenotypes between L. major and T. brucei, in contrast to the phenotypes associated with the epigenetic DNA base J modification. Total RNA from Four LmjF samples were analyzed using RNA-Seq. One of them is wildtype parasites, one is single knockout for H3V gene and two independent double knockouts for H3V gene.

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Deep proteome analysis of Leishmania major Friedlin

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