Identification and characterization of nuclear and nucleolar localization signals in 58-kDa microspherule protein (MSP58)

Chuan Pin Yang, Chi Wu Chiang, Chang Han Chen, Yi Chao Lee, Mei Hsiang Wu, Yi Huan Tsou, Yu San Yang, Wen Chang Chang, Ding Yen Lin

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Background: MSP58 is a nucleolar protein associated with rRNA transcription and cell proliferation. Its mechanism of translocation into the nucleus or the nucleolus, however, is not entirely known. In order to address this lack, the present study aims to determine a crucial part of this mechanism: the nuclear localization signal (NLS) and the nucleolar localization signal (NoLS) associated with the MSP58 protein. Results: We have identified and characterized two NLSs in MSP58. The first is located between residues 32 and 56 (NLS1) and constitutes three clusters of basic amino acids (KRASSQALGTIPKRRSSSRFIKRKK); the second is situated between residues 113 and 123 (NLS2) and harbors a monopartite signal (PGLTKRVKKSK). Both NLS1 and NLS2 are highly conserved among different vertebrate species. Notably, one bipartite motif within the NLS1 (residues 44-56) appears to be absolutely necessary for MSP58 nucleolar localization. By yeast two-hybrid, pull-down, and coimmunoprecipitation analysis, we show that MSP58 binds to importin α1 and α6, suggesting that nuclear targeting of MSP58 utilizes a receptor-mediated and energy-dependent import mechanism. Functionally, our data show that both nuclear and nucleolar localization of MSP58 are crucial for transcriptional regulation on p21 and ribosomal RNA genes, and context-dependent effects on cell proliferation. Conclusions: Results suggest that MSP58 subnuclear localization is regulated by two nuclear import signals, and that proper subcellular localization of MSP58 is critical for its role in transcriptional regulation. Our study reveals a molecular mechanism that controls nuclear and nucleolar localization of MSP58, a finding that might help future researchers understand the MSP58 biological signaling pathway.

Original languageEnglish
Article number33
JournalJournal of Biomedical Science
Volume22
Issue number1
DOIs
Publication statusPublished - May 16 2015

Fingerprint

Nuclear Localization Signals
Cell proliferation
Cell Proliferation
Karyopherins
Basic Amino Acids
Ribosomal RNA
Cell Nucleus Active Transport
Transcription
Nuclear Proteins
Ports and harbors
rRNA Genes
Yeast
Vertebrates
Proteins
Genes
Yeasts
Research Personnel

Keywords

  • 58-kDa Microspherule Protein
  • Importins
  • Nuclear localization signal
  • Nucleolar localization signal
  • Nucleolus

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Molecular Biology
  • Cell Biology
  • Biochemistry, medical
  • Endocrinology, Diabetes and Metabolism
  • Pharmacology (medical)

Cite this

Identification and characterization of nuclear and nucleolar localization signals in 58-kDa microspherule protein (MSP58). / Yang, Chuan Pin; Chiang, Chi Wu; Chen, Chang Han; Lee, Yi Chao; Wu, Mei Hsiang; Tsou, Yi Huan; Yang, Yu San; Chang, Wen Chang; Lin, Ding Yen.

In: Journal of Biomedical Science, Vol. 22, No. 1, 33, 16.05.2015.

Research output: Contribution to journalArticle

Yang, Chuan Pin ; Chiang, Chi Wu ; Chen, Chang Han ; Lee, Yi Chao ; Wu, Mei Hsiang ; Tsou, Yi Huan ; Yang, Yu San ; Chang, Wen Chang ; Lin, Ding Yen. / Identification and characterization of nuclear and nucleolar localization signals in 58-kDa microspherule protein (MSP58). In: Journal of Biomedical Science. 2015 ; Vol. 22, No. 1.
@article{320f59604acf4a4e9207242347715a6f,
title = "Identification and characterization of nuclear and nucleolar localization signals in 58-kDa microspherule protein (MSP58)",
abstract = "Background: MSP58 is a nucleolar protein associated with rRNA transcription and cell proliferation. Its mechanism of translocation into the nucleus or the nucleolus, however, is not entirely known. In order to address this lack, the present study aims to determine a crucial part of this mechanism: the nuclear localization signal (NLS) and the nucleolar localization signal (NoLS) associated with the MSP58 protein. Results: We have identified and characterized two NLSs in MSP58. The first is located between residues 32 and 56 (NLS1) and constitutes three clusters of basic amino acids (KRASSQALGTIPKRRSSSRFIKRKK); the second is situated between residues 113 and 123 (NLS2) and harbors a monopartite signal (PGLTKRVKKSK). Both NLS1 and NLS2 are highly conserved among different vertebrate species. Notably, one bipartite motif within the NLS1 (residues 44-56) appears to be absolutely necessary for MSP58 nucleolar localization. By yeast two-hybrid, pull-down, and coimmunoprecipitation analysis, we show that MSP58 binds to importin α1 and α6, suggesting that nuclear targeting of MSP58 utilizes a receptor-mediated and energy-dependent import mechanism. Functionally, our data show that both nuclear and nucleolar localization of MSP58 are crucial for transcriptional regulation on p21 and ribosomal RNA genes, and context-dependent effects on cell proliferation. Conclusions: Results suggest that MSP58 subnuclear localization is regulated by two nuclear import signals, and that proper subcellular localization of MSP58 is critical for its role in transcriptional regulation. Our study reveals a molecular mechanism that controls nuclear and nucleolar localization of MSP58, a finding that might help future researchers understand the MSP58 biological signaling pathway.",
keywords = "58-kDa Microspherule Protein, Importins, Nuclear localization signal, Nucleolar localization signal, Nucleolus",
author = "Yang, {Chuan Pin} and Chiang, {Chi Wu} and Chen, {Chang Han} and Lee, {Yi Chao} and Wu, {Mei Hsiang} and Tsou, {Yi Huan} and Yang, {Yu San} and Chang, {Wen Chang} and Lin, {Ding Yen}",
year = "2015",
month = "5",
day = "16",
doi = "10.1186/s12929-015-0136-0",
language = "English",
volume = "22",
journal = "Journal of Biomedical Science",
issn = "1021-7770",
publisher = "BioMed Central",
number = "1",

}

TY - JOUR

T1 - Identification and characterization of nuclear and nucleolar localization signals in 58-kDa microspherule protein (MSP58)

AU - Yang, Chuan Pin

AU - Chiang, Chi Wu

AU - Chen, Chang Han

AU - Lee, Yi Chao

AU - Wu, Mei Hsiang

AU - Tsou, Yi Huan

AU - Yang, Yu San

AU - Chang, Wen Chang

AU - Lin, Ding Yen

PY - 2015/5/16

Y1 - 2015/5/16

N2 - Background: MSP58 is a nucleolar protein associated with rRNA transcription and cell proliferation. Its mechanism of translocation into the nucleus or the nucleolus, however, is not entirely known. In order to address this lack, the present study aims to determine a crucial part of this mechanism: the nuclear localization signal (NLS) and the nucleolar localization signal (NoLS) associated with the MSP58 protein. Results: We have identified and characterized two NLSs in MSP58. The first is located between residues 32 and 56 (NLS1) and constitutes three clusters of basic amino acids (KRASSQALGTIPKRRSSSRFIKRKK); the second is situated between residues 113 and 123 (NLS2) and harbors a monopartite signal (PGLTKRVKKSK). Both NLS1 and NLS2 are highly conserved among different vertebrate species. Notably, one bipartite motif within the NLS1 (residues 44-56) appears to be absolutely necessary for MSP58 nucleolar localization. By yeast two-hybrid, pull-down, and coimmunoprecipitation analysis, we show that MSP58 binds to importin α1 and α6, suggesting that nuclear targeting of MSP58 utilizes a receptor-mediated and energy-dependent import mechanism. Functionally, our data show that both nuclear and nucleolar localization of MSP58 are crucial for transcriptional regulation on p21 and ribosomal RNA genes, and context-dependent effects on cell proliferation. Conclusions: Results suggest that MSP58 subnuclear localization is regulated by two nuclear import signals, and that proper subcellular localization of MSP58 is critical for its role in transcriptional regulation. Our study reveals a molecular mechanism that controls nuclear and nucleolar localization of MSP58, a finding that might help future researchers understand the MSP58 biological signaling pathway.

AB - Background: MSP58 is a nucleolar protein associated with rRNA transcription and cell proliferation. Its mechanism of translocation into the nucleus or the nucleolus, however, is not entirely known. In order to address this lack, the present study aims to determine a crucial part of this mechanism: the nuclear localization signal (NLS) and the nucleolar localization signal (NoLS) associated with the MSP58 protein. Results: We have identified and characterized two NLSs in MSP58. The first is located between residues 32 and 56 (NLS1) and constitutes three clusters of basic amino acids (KRASSQALGTIPKRRSSSRFIKRKK); the second is situated between residues 113 and 123 (NLS2) and harbors a monopartite signal (PGLTKRVKKSK). Both NLS1 and NLS2 are highly conserved among different vertebrate species. Notably, one bipartite motif within the NLS1 (residues 44-56) appears to be absolutely necessary for MSP58 nucleolar localization. By yeast two-hybrid, pull-down, and coimmunoprecipitation analysis, we show that MSP58 binds to importin α1 and α6, suggesting that nuclear targeting of MSP58 utilizes a receptor-mediated and energy-dependent import mechanism. Functionally, our data show that both nuclear and nucleolar localization of MSP58 are crucial for transcriptional regulation on p21 and ribosomal RNA genes, and context-dependent effects on cell proliferation. Conclusions: Results suggest that MSP58 subnuclear localization is regulated by two nuclear import signals, and that proper subcellular localization of MSP58 is critical for its role in transcriptional regulation. Our study reveals a molecular mechanism that controls nuclear and nucleolar localization of MSP58, a finding that might help future researchers understand the MSP58 biological signaling pathway.

KW - 58-kDa Microspherule Protein

KW - Importins

KW - Nuclear localization signal

KW - Nucleolar localization signal

KW - Nucleolus

UR - http://www.scopus.com/inward/record.url?scp=84933504842&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84933504842&partnerID=8YFLogxK

U2 - 10.1186/s12929-015-0136-0

DO - 10.1186/s12929-015-0136-0

M3 - Article

C2 - 25981436

AN - SCOPUS:84933504842

VL - 22

JO - Journal of Biomedical Science

JF - Journal of Biomedical Science

SN - 1021-7770

IS - 1

M1 - 33

ER -