Dataset Information

Functional analyses of human LUC7-like proteins involved in splicing regulation and myeloid neoplasms

ABSTRACT: We chracaterized the functions of the understudied LUC7-like family of splicing factors in human cells using seCLIP-seq, RBP knockdown followed by RNA-seq, and Co-IP mass spectrometry.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Cell, Cell Suspension Culture

DISEASE(S): Acute Leukemia

SUBMITTER: Noah Daniels

LAB HEAD: Richard A. Padgett

PROVIDER: PXD022152 | Pride | 2021-09-09

REPOSITORIES: Pride

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Publications

Functional analyses of human LUC7-like proteins involved in splicing regulation and myeloid neoplasms.

Daniels Noah J NJ Hershberger Courtney E CE Gu Xiaorong X Schueger Caroline C DiPasquale William M WM Brick Jonathan J Saunthararajah Yogen Y Maciejewski Jaroslaw P JP Padgett Richard A RA

Cell reports 20210401 2

Vertebrates have evolved three paralogs, termed LUC7L, LUC7L2, and LUC7L3, of the essential yeast U1 small nuclear RNA (snRNA)-associated splicing factor Luc7p. We investigated the mechanistic and regulatory functions of these putative splicing factors, of which one (LUC7L2) is mutated or deleted in myeloid neoplasms. Protein interaction data show that all three proteins bind similar core but distinct regulatory splicing factors, probably mediated through their divergent arginine-serine-rich dom ...[more]

PMID: 33852859

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Project description:We aim to understand the role that Cdx2 plays in specifying the rostro-caudal identity of differentiating motor neurons. We find that expressing Cdx2 in combination with FGF signaling is sufficient to produce motor neurons with a more caudal identity. ChIP-seq analysis of Cdx2 finds that it binds extensively throughout the Hox regions in progenitor motor neurons. Analysis of polycomb-associated chromatin over Hox regions in the subsequently generated motor neurons finds that Cdx2 binding corresponds to chromatin domains encompassing de-repressed caudal Hox genes. These results suggest a direct role for Cdx2 in specifying caudal motor neuron identity. ChIP-seq studies: We characterize the binding of Cdx2 in progenitor motor neurons using a V5 tagged doxycycline inducible Cdx2 ESC line (iCdx2). Progenitor motor neurons were generated after 4 days of in vitro differentiation of mouse embryonic stem cells using retinoic acid (RA) and hedgehog (Hh) signaling exposure at day 2. On day 3, the cells are exposed to Dox with and without accompanying FGF signaling. The genome-wide binding of the induced Cdx2 transcription factor is profiled using ChIP-seq with an anti-V5 antibody. An appropriate whole-cell extract control experiment for these ChIP-seq experiments is also included. We also examine the effect of induced Cdx2 expression on polycomb-associated chromatin structure in the resulting cellular populations by profiling the H3K27me3 chromatin mark using ChIP-seq. H3K27me3 experiments were performed after 5 days of in vitro differentiation using cells exposed to either: 1) RA & Hh to derive progenitor motor neurons, followed by Dox & FGF; 2) Dox & FGF alone; or 3) RA and Hh alone. There are 6 Illumina sequencing datasets included in this submission: two biological replicates of iCdx2 ChIP-seq in the presence of FGF; one sample of iCdx2 ChIP-seq in the absence of FGF; one H3K27me3 ChIP-seq in the presence of RA, Hh, Dox, and FGF; one H3K27me3 ChIP-seq in the presence of Dox and FGF; and one H3K27me3 ChIP-seq in the presence of RA and Hh.

Dataset Information

Functional analyses of human LUC7-like proteins involved in splicing regulation and myeloid neoplasms

Publications

Functional analyses of human LUC7-like proteins involved in splicing regulation and myeloid neoplasms.

Similar Datasets

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