Universität zu Köln

Burgute, Bhagyashri D., Peche, Vivek S., Steckelberg, Anna-Lena ORCID: 0000-0001-7988-9946, Gloeckner, Gernot, Gassen, Berthold, Gehring, Niels H. ORCID: 0000-0001-7792-1164 and Noegel, Angelika A. (2014). NKAP is a novel RS-related protein that interacts with RNA and RNA binding proteins. Nucleic Acids Res., 42 (5). S. 3177 - 3194. OXFORD: OXFORD UNIV PRESS. ISSN 1362-4962

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Abstract

NKAP is a highly conserved protein with roles in transcriptional repression, T-cell development, maturation and acquisition of functional competency and maintenance and survival of adult hematopoietic stem cells. Here we report the novel role of NKAP in splicing. With NKAP-specific antibodies we found that NKAP localizes to nuclear speckles. NKAP has an RS motif at the N-terminus followed by a highly basic domain and a DUF 926 domain at the C-terminal region. Deletion analysis showed that the basic domain is important for speckle localization. In pull-down experiments, we identified RNA-binding proteins, RNA helicases and splicing factors as interaction partners of NKAP, among them FUS/TLS. The FUS/TLS-NKAP interaction takes place through the RS domain of NKAP and the RGG1 and RGG3 domains of FUS/TLS. We analyzed the ability of NKAP to interact with RNA using in vitro splicing assays and found that NKAP bound both spliced messenger RNA (mRNA) and unspliced pre-mRNA. Genome-wide analysis using crosslinking and immunoprecipitation-seq revealed NKAP association with U1, U4 and U5 small nuclear RNA, and we also demonstrated that knockdown of NKAP led to an increase in pre-mRNA percentage. Our results reveal NKAP as nuclear speckle protein with roles in RNA splicing and processing.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Burgute, Bhagyashri D. UNSPECIFIED UNSPECIFIED UNSPECIFIED Peche, Vivek S. UNSPECIFIED UNSPECIFIED UNSPECIFIED Steckelberg, Anna-Lena UNSPECIFIED orcid.org/0000-0001-7988-9946 UNSPECIFIED Gloeckner, Gernot UNSPECIFIED UNSPECIFIED UNSPECIFIED Gassen, Berthold UNSPECIFIED UNSPECIFIED UNSPECIFIED Gehring, Niels H. UNSPECIFIED orcid.org/0000-0001-7792-1164 UNSPECIFIED Noegel, Angelika A. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-444576
DOI:	10.1093/nar/gkt1311
Journal or Publication Title:	Nucleic Acids Res.
Volume:	42
Number:	5
Page Range:	S. 3177 - 3194
Date:	2014
Publisher:	OXFORD UNIV PRESS
Place of Publication:	OXFORD
ISSN:	1362-4962
Language:	English
Faculty:	Faculty of Mathematics and Natural Sciences
Divisions:	Faculty of Mathematics and Natural Sciences > Department of Biology > Institute for Genetics
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language SPLICING FACTOR SC35; SERINE-RICH DOMAINS; MESSENGER-RNA; NUCLEAR; IDENTIFICATION; GENE; LOCALIZATION; PURIFICATION; TLS; SPLICEOSOME Multiple languages Biochemistry & Molecular Biology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/44457