NIPP1 (351 residues) is a major regulatory and RNA-anchoring subunit of protein phosphatase 1 in the nucleus. Using recombinant and synthetic fragments of NIPP1, the RNA-binding domain was mapped to the C-terminal residues 330-351. A synthetic peptide encompassing this sequence equalled intact NIPP1 in RNA-binding affinity and could be used to dissociate NIPP1 from the nuclear particulate fraction. An NIPP1 fragment consisting of residues 225-351 (Ard1/NIPP1γ), that may be encoded by an alternatively spliced transcript in transformed B-lymphocytes, displayed a single-strand Mg2+-dependent endoribonuclease activity. However, full-length NIPP1 and NIPP1143-351 were not able to cleave RNA, indicating that the endoribonuclease activity of NIPP1 is restrained by its central domain. The endoribonuclease activity was also recovered in the RNA-binding domain, NIPP1330-351, but with a 30-fold lower specific activity. Thus, the endoribonuclease catalytic site and the RNA-binding site both reside in the C-terminal 22 residues of NIPP1. The latter domain does not conform to any known nucleic-acid binding motif.
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Research Article|
August 10 1999
Mapping of the RNA-binding and endoribonuclease domains of NIPP1, a nuclear targeting subunit of protein phosphatase 1
Qiming JIN;
Qiming JIN
1Afdeling Biochemie, Faculteit Geneeskunde, Katholieke Universiteit Leuven, B-3000 Leuven, Belgium
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Monique BEULLENS;
Monique BEULLENS
1Afdeling Biochemie, Faculteit Geneeskunde, Katholieke Universiteit Leuven, B-3000 Leuven, Belgium
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Izabela JAGIELLO;
Izabela JAGIELLO
1Afdeling Biochemie, Faculteit Geneeskunde, Katholieke Universiteit Leuven, B-3000 Leuven, Belgium
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Aleyde VAN EYNDE;
Aleyde VAN EYNDE
1Afdeling Biochemie, Faculteit Geneeskunde, Katholieke Universiteit Leuven, B-3000 Leuven, Belgium
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Veerle VULSTEKE;
Veerle VULSTEKE
1Afdeling Biochemie, Faculteit Geneeskunde, Katholieke Universiteit Leuven, B-3000 Leuven, Belgium
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Willy STALMANS;
Willy STALMANS
1Afdeling Biochemie, Faculteit Geneeskunde, Katholieke Universiteit Leuven, B-3000 Leuven, Belgium
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Mathieu BOLLEN
Mathieu BOLLEN
1
1Afdeling Biochemie, Faculteit Geneeskunde, Katholieke Universiteit Leuven, B-3000 Leuven, Belgium
1To whom correspondence should be addressed, at Afdeling Biochemie, Campus Gasthuisberg, Herestraat 49, B-3000 Leuven, Belgium (e-mail Mathieu.Bollen@med.kuleuven.ac.be).
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Publisher: Portland Press Ltd
Received:
March 23 1999
Revision Received:
May 10 1999
Accepted:
June 14 1999
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1999
1999
Biochem J (1999) 342 (1): 13–19.
Article history
Received:
March 23 1999
Revision Received:
May 10 1999
Accepted:
June 14 1999
Citation
Qiming JIN, Monique BEULLENS, Izabela JAGIELLO, Aleyde VAN EYNDE, Veerle VULSTEKE, Willy STALMANS, Mathieu BOLLEN; Mapping of the RNA-binding and endoribonuclease domains of NIPP1, a nuclear targeting subunit of protein phosphatase 1. Biochem J 15 August 1999; 342 (1): 13–19. doi: https://doi.org/10.1042/bj3420013
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