Dataset Information

Elucidation of the role of murine lncRNA LL35 in hepatocytes by LC-MSMS

ABSTRACT: Long non-coding RNAs (lncRNAs) play important roles in the regulation of many biological processes in the cell like transcription, translation, splicing, transport, etc. More than 10K lncRNAs are predicted in human, but functions for the majority remain unknown. LncRNAs may contribute to the development of multiple diseases that makes them perspective diagnostic and prognostic markers as well as drug targets [1]. Murine lncRNA Falcor/LL35 is a proposed functional analog of human lncRNA DEANR1, which is predominantly expressed in murine lungs and liver, intestine and pancreas [2, 3]. While the participation of LL35 in LL35–Foxa2 (transcription factor) regulatory loop during regeneration after lung injury was previously described by Swarr et al. [2], its functional role in murine liver remains unknown. In our work we focused on revealing the role of murine lncRNA LL35 in hepatocytes and in mouse liver. We analyzed changes in proteome of AML12 (normal hepatocytes) cell line on the 2nd day after LL35 depletion. [1] Morlando, M., Ballarino, M., & Fatica, A. (2015). Long non-coding RNAs: New players in hematopoiesis and leukemia. Frontiers in Medicine, Vol. 2. [2] Swarr, D. T., Herriges, M., Li, S., Morley, M., Fernandes, S., Sridharan, A., … Morrisey, E. E. (2019). The long noncoding RNA Falcor regulates Foxa2 expression to maintain lung epithelial homeostasis and promote regeneration. Genes & Development, 33(11-12), 656–668. [3] Sergeeva, O. V, Korinfskaya, S. A., Kurochkin, I. I., & Zatsepin, T. S. (2019). Long Noncoding RNA LL35 / Falcor Regulates Expression of Transcription Factor Foxa2 in Hepatocytes in Normal and Fibrotic Mouse Liver. Acta Naturae, 11(42), 66–74.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Hepatocyte, Cell Culture

SUBMITTER: Rustam Ziganshin

LAB HEAD: Timofei S. Zatsepin

PROVIDER: PXD033910 | Pride | 2022-08-11

REPOSITORIES: Pride

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			Action	DRS
	AF_031_20200224_ZR_AF-1-1.raw	Raw
	AF_032_20200224_ZR_AF-1-2.raw	Raw
	AF_033_20200224_ZR_AF-1-3.raw	Raw
	AF_035_20200224_ZR_AF-1-4.raw	Raw
	AF_036_20200224_ZR_AF-1-5.raw	Raw

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Publications

Murine Falcor/LL35 lncRNA Contributes to Glucose and Lipid Metabolism In Vitro and In Vivo.

Shcherbinina Evgeniya E Abakumova Tatiana T Bobrovskiy Daniil D Kurochkin Ilia I Deinichenko Ksenia K Stekolshchikova Elena E Anikanov Nickolay N Ziganshin Rustam R Melnikov Pavel P Khrameeva Ekaterina E Logacheva Maria M Zatsepin Timofei T Sergeeva Olga O

Biomedicines 20220613 6

Glucose and lipid metabolism are crucial functional systems in eukaryotes. A large number of experimental studies both in animal models and humans have shown that long non-coding RNAs (lncRNAs) play an important role in glucose and lipid metabolism. Previously, human lncRNA DEANR1/linc00261 was described as a tumor suppressor that regulates a variety of biological processes such as cell proliferation, apoptosis, glucose metabolism and tumorigenesis. Here we report that murine lncRNA Falcor/LL35, ...[more]

PMID: 35740417

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Project description:Mammals express thousands of long noncoding (lnc) RNAs, a few of which are shown to function in tissue development. However, the entire repertoire of lncRNAs and the extent to which they regulate biological processes in different tissues and species are not defined. Indeed, most lncRNAs are not conserved between species, raising questions about function. We used RNA-Seq to identify lncRNAs in primary murine fetal liver erythroblasts expressing the lineage marker TER119, megakaryocytes (CD41+) cultured from embryonic day (E) 14.5 murine fetal liver and megakaryocyte erythroid progenitors (MEPs) isolated from mouse bone marrow. We identified 683 and 594 polyadenylated lncRNAs expressed in red blood cell (erythroid) precursors of mice and humans, respectively. More than one half of erythroid lncRNAs are un-annotated, emphasizing the opportunity for new discovery through studies of specialized cell types. We analyzed the expression of these identified lncRNAs in several hematopoietic compartments using a custom microarray to identify erythroid-specific lncRNAs that were robustly expressed in both fetal liver and adult erythroid cells as targets for knockdown. Over 90% of fetal liver erythroid lncRNAs detected using RNA-seq were expressed in adult erythroblasts measured on the microarray. Analysis of the murine erythroid lncRNA transcriptome indicates that ~75% arise from promoters and 25% from enhancers, many of which are regulated by the key erythroid transcription factors GATA1 and SCL/TAL1. Erythroid lncRNA expression is largely conserved among 8 different mouse strains, yet only 15% of mouse lncRNAs are expressed in humans and vice versa, reflecting dramatically greater species-specificity than coding genes. We investigated potential functions of 21 relatively abundant erythroid-specific murine lncRNAs (both conserved and non-conserved) by RNA interference in primary mouse erythroid precursors, and identified 7 whose knockdown inhibited features of terminal erythroid maturation including cell size reduction and enucleation. Strikingly, at least 6 of the 7 lncRNAs have no detectable expression in human erythroblasts, demonstrating that lack of conservation between mammalian species does not predict lack of function. These results reflect marked evolutionary differences between protein-coding genes and lncRNAs and indicate that the latter exert tissue- and species-specific roles in development. For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf A custom Agilent microarray was designed to interrogate expression levels of long noncoding RNAs identified using RNA-seq in several different hematopoietic progenitor and differentiated cell populations. LncRNA gene definitions and RNA-seq data used to identify long noncoding RNAs are deposited in GEO with the accession numbers GSE51667 and GSE40522 respectively. The Agilent eArray platform was used to customize the SurePrint G3 Mouse GE 8x60K microarray to add 3 custom-designed probes (60nt length) each against several categories of genes, namely pseudogene, genes with small RNA overlap, low stringency lncRNAs and high stringency lncRNAs . We interrogated 7 types of hematopoietic cells, HSC, CMP, MEP, GMP, early and late erythroblasts and granulocytes. Expression measurements were determined at least in duplicate for all samples and in triplicate for most samples.