Burgute, Bhagyashri D., Peche, Vivek S., Steckelberg, Anna-Lena ORCID: 0000-0001-7988-9946, Gloeckner, Gernot, Gassen, Berthold, Gehring, Niels H. 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:
CreatorsEmailORCIDORCID Put Code
Burgute, Bhagyashri D.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Peche, Vivek S.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Steckelberg, Anna-LenaUNSPECIFIEDorcid.org/0000-0001-7988-9946UNSPECIFIED
Gloeckner, GernotUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Gassen, BertholdUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Gehring, Niels H.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Noegel, Angelika A.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
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: Unspecified
Divisions: Unspecified
Subjects: no entry
Uncontrolled Keywords:
KeywordsLanguage
SPLICING FACTOR SC35; SERINE-RICH DOMAINS; MESSENGER-RNA; NUCLEAR; IDENTIFICATION; GENE; LOCALIZATION; PURIFICATION; TLS; SPLICEOSOMEMultiple languages
Biochemistry & Molecular BiologyMultiple languages
URI: http://kups.ub.uni-koeln.de/id/eprint/44457

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