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Identifying lncRNA-protein interactions in hematopoietic progenitor cells by hybridization capture and mass spectrometry

ABSTRACT: Long non-coding RNAs (lncRNAs) exert regulatory functions in a wide spectrum of biological contexts, and certain regulatory functions involve the formation of RNA-protein complexes. Discovering the structure/function of these complexes may unveil important functional insights. The DDX41 gene encoding the DEAD-box RNA helicase 41 protein (DDX41) is subject to extensive germline genetic variation, and certain variants create a predisposition to develop myelodysplastic syndrome and acute myeloid leukemia. While the importance of DDX41 for the control of hematopoiesis is established, many questions remain regarding the mechanisms of how DDX41 functions in hematopoietic stem and progenitor cells. Previously, we identified a DDX41-regulated lncRNA, growth arrest-specific 5 (Gas5). As Gas5 function in hematopoiesis is unknown, we analyzed protein interactors of Gas5 lncRNA using HyPR-MS (Hybridization Purification of RNA-Protein complexes followed by Mass Spectrometry). 303 proteins were identified as Gas5 lncRNA interactors, five of which were experimentally validated as Gas5 lncRNA interactors by RNA Immunoprecipitation qPCR (RIP-qPCR) analysis. The identification of protein interactors with a DDX41-regulated lncRNA establishes a foundation to guide future mechanistic and biological studies.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Stem Cell, Cell Culture

SUBMITTER: Yuling Dai

LAB HEAD: Lloyd M.

PROVIDER: PXD062695 | Pride | 2025-07-17

REPOSITORIES: Pride

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			Action	DRS
	AllPSMs.psmtsv	Other
	AllPSMs_FormattedForPercolator.tab	Other
	AllPeptides.psmtsv	Other
	AllQuantifiedPeptides.tsv	Tabular
	AllQuantifiedProteinGroups.tsv	Tabular

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lncRNA-protein interactions in hematopoietic progenitor cells by hybridization capture and mass spectrometry

Project description:Long non-coding RNAs (lncRNAs) exert regulatory functions in a wide spectrum of biological contexts, and certain regulatory functions involve the formation of RNA-protein complexes. Discovering the structure/function of these complexes may unveil important functional insights. The DDX41 gene encoding the DEAD-box RNA helicase 41 protein (DDX41) is subject to extensive germline genetic variation, and certain variants create a predisposition to develop myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). While the importance of DDX41 for the control of hematopoiesis is established, many questions remain regarding the mechanisms of how DDX41 functions in hematopoietic stem and progenitor cells. Previously, we identified a DDX41-regulated lncRNA, growth arrest-specific 5 (Gas5) (Kim et al.). As Gas5 function in hematopoiesis is unknown, we analyzed protein interactors of Gas5 lncRNA using HyPR-MS (Hybridization Purification of RNA-Protein complexes followed by Mass Spectrometry). 28 proteins were identified as Gas5 lncRNA interactors, five of which were experimentally validated as Gas5 lncRNA interactors by RNA Immunoprecipitation qPCR (RIP-qPCR) analysis. The identification of protein interactors with a DDX41-regulated lncRNA establishes a foundation to guide future mechanistic and biological studies.c

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