Identification of a nuclear localization signal mediating the nuclear import of Arabidopsis splicing factor1

Eun Jin Wang; Young Cheon Kim; Jeong Hwan Lee; Jeong Kook Kim

doi:10.1007/s11816-021-00722-0

Identification of a nuclear localization signal mediating the nuclear import of Arabidopsis splicing factor1

Eun Jin Wang, Young Cheon Kim, Jeong Hwan Lee, Jeong Kook Kim

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Abstract

The splicing factor1 protein (SF1) is involved in branch point recognition of pre-mRNA introns during the early stages of spliceosome assembly in the nucleus. In this study, we aimed to characterize the nuclear localization signal (NLS) of the Arabidopsis SF1 protein (AtSF1). There are two putative NLS sequences (RRKRRSR and RKRKSR) at the N-terminal side of the AtSF1 protein. Analysis of green fluorescence protein (GFP)-tagged AtSF1 deletion constructs indicated that the RKRKSRWADDE sequence (from the 124th to 134th amino acid residues) is necessary for GFP-tagged AtSF1 protein for the localization in the nucleus. Further analysis of the RKRKSRWADDE sequence using site-directed mutagenesis demonstrated that at least two basic amino acid residues (R and K) within the sequence is essential for the complete nuclear localization of GFP-tagged AtSF1 protein. Taken together, our findings demonstrated that only one of the two predicted NLS candidates of the AtSF1 protein is necessary for its nuclear localization, and at least two basic amino acid residues within the motif are crucial for its function. This feature of NLS may be unique in plant SF1 proteins because there is only one predicted NLS in fungal and metazoan counterparts.

Original language	English
Pages (from-to)	775-781
Number of pages	7
Journal	Plant Biotechnology Reports
Volume	15
Issue number	6
DOIs	https://doi.org/10.1007/s11816-021-00722-0
Publication status	Published - 2021 Dec

Bibliographical note

Funding Information:
Financial support for this work was provided by the Basic Science Research Program through a National Research Foundation of Korea (NRF) grant funded by the Ministry of Education (2019R1F1A1060009 to J.-K. Kim), and a Korea University Grant (to J.-K. Kim). This work was also supported in part by a grant (PJ01532503 to J.H. Lee) from the Rural Development Administration (RDA), Republic of Korea.

Funding Information:
Financial support for this work was provided by the Basic Science Research Program through a National Research Foundation of Korea (NRF) grant funded by the Ministry of Education (2019R1F1A1060009 to J.-K. Kim), and a Korea University Grant (to J.-K. Kim). This work was also supported in part by a grant (PJ01532503 to J.H. Lee) from the Rural Development Administration (RDA), Republic of Korea.

Publisher Copyright:
© 2021, Korean Society for Plant Biotechnology.

Keywords

Nuclear localization signal
Nucleocytoplasmic shuttling
Splicing factor1 protein
U2AF ligand motif

ASJC Scopus subject areas

Biotechnology
Plant Science

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10.1007/s11816-021-00722-0

Cite this

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title = "Identification of a nuclear localization signal mediating the nuclear import of Arabidopsis splicing factor1",

abstract = "The splicing factor1 protein (SF1) is involved in branch point recognition of pre-mRNA introns during the early stages of spliceosome assembly in the nucleus. In this study, we aimed to characterize the nuclear localization signal (NLS) of the Arabidopsis SF1 protein (AtSF1). There are two putative NLS sequences (RRKRRSR and RKRKSR) at the N-terminal side of the AtSF1 protein. Analysis of green fluorescence protein (GFP)-tagged AtSF1 deletion constructs indicated that the RKRKSRWADDE sequence (from the 124th to 134th amino acid residues) is necessary for GFP-tagged AtSF1 protein for the localization in the nucleus. Further analysis of the RKRKSRWADDE sequence using site-directed mutagenesis demonstrated that at least two basic amino acid residues (R and K) within the sequence is essential for the complete nuclear localization of GFP-tagged AtSF1 protein. Taken together, our findings demonstrated that only one of the two predicted NLS candidates of the AtSF1 protein is necessary for its nuclear localization, and at least two basic amino acid residues within the motif are crucial for its function. This feature of NLS may be unique in plant SF1 proteins because there is only one predicted NLS in fungal and metazoan counterparts.",

keywords = "Nuclear localization signal, Nucleocytoplasmic shuttling, Splicing factor1 protein, U2AF ligand motif",

author = "Wang, {Eun Jin} and Kim, {Young Cheon} and Lee, {Jeong Hwan} and Kim, {Jeong Kook}",

note = "Funding Information: Financial support for this work was provided by the Basic Science Research Program through a National Research Foundation of Korea (NRF) grant funded by the Ministry of Education (2019R1F1A1060009 to J.-K. Kim), and a Korea University Grant (to J.-K. Kim). This work was also supported in part by a grant (PJ01532503 to J.H. Lee) from the Rural Development Administration (RDA), Republic of Korea. Funding Information: Financial support for this work was provided by the Basic Science Research Program through a National Research Foundation of Korea (NRF) grant funded by the Ministry of Education (2019R1F1A1060009 to J.-K. Kim), and a Korea University Grant (to J.-K. Kim). This work was also supported in part by a grant (PJ01532503 to J.H. Lee) from the Rural Development Administration (RDA), Republic of Korea. Publisher Copyright: {\textcopyright} 2021, Korean Society for Plant Biotechnology.",

year = "2021",

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doi = "10.1007/s11816-021-00722-0",

language = "English",

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AU - Kim, Young Cheon

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AU - Kim, Jeong Kook

N1 - Funding Information: Financial support for this work was provided by the Basic Science Research Program through a National Research Foundation of Korea (NRF) grant funded by the Ministry of Education (2019R1F1A1060009 to J.-K. Kim), and a Korea University Grant (to J.-K. Kim). This work was also supported in part by a grant (PJ01532503 to J.H. Lee) from the Rural Development Administration (RDA), Republic of Korea. Funding Information: Financial support for this work was provided by the Basic Science Research Program through a National Research Foundation of Korea (NRF) grant funded by the Ministry of Education (2019R1F1A1060009 to J.-K. Kim), and a Korea University Grant (to J.-K. Kim). This work was also supported in part by a grant (PJ01532503 to J.H. Lee) from the Rural Development Administration (RDA), Republic of Korea. Publisher Copyright: © 2021, Korean Society for Plant Biotechnology.

PY - 2021/12

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N2 - The splicing factor1 protein (SF1) is involved in branch point recognition of pre-mRNA introns during the early stages of spliceosome assembly in the nucleus. In this study, we aimed to characterize the nuclear localization signal (NLS) of the Arabidopsis SF1 protein (AtSF1). There are two putative NLS sequences (RRKRRSR and RKRKSR) at the N-terminal side of the AtSF1 protein. Analysis of green fluorescence protein (GFP)-tagged AtSF1 deletion constructs indicated that the RKRKSRWADDE sequence (from the 124th to 134th amino acid residues) is necessary for GFP-tagged AtSF1 protein for the localization in the nucleus. Further analysis of the RKRKSRWADDE sequence using site-directed mutagenesis demonstrated that at least two basic amino acid residues (R and K) within the sequence is essential for the complete nuclear localization of GFP-tagged AtSF1 protein. Taken together, our findings demonstrated that only one of the two predicted NLS candidates of the AtSF1 protein is necessary for its nuclear localization, and at least two basic amino acid residues within the motif are crucial for its function. This feature of NLS may be unique in plant SF1 proteins because there is only one predicted NLS in fungal and metazoan counterparts.

AB - The splicing factor1 protein (SF1) is involved in branch point recognition of pre-mRNA introns during the early stages of spliceosome assembly in the nucleus. In this study, we aimed to characterize the nuclear localization signal (NLS) of the Arabidopsis SF1 protein (AtSF1). There are two putative NLS sequences (RRKRRSR and RKRKSR) at the N-terminal side of the AtSF1 protein. Analysis of green fluorescence protein (GFP)-tagged AtSF1 deletion constructs indicated that the RKRKSRWADDE sequence (from the 124th to 134th amino acid residues) is necessary for GFP-tagged AtSF1 protein for the localization in the nucleus. Further analysis of the RKRKSRWADDE sequence using site-directed mutagenesis demonstrated that at least two basic amino acid residues (R and K) within the sequence is essential for the complete nuclear localization of GFP-tagged AtSF1 protein. Taken together, our findings demonstrated that only one of the two predicted NLS candidates of the AtSF1 protein is necessary for its nuclear localization, and at least two basic amino acid residues within the motif are crucial for its function. This feature of NLS may be unique in plant SF1 proteins because there is only one predicted NLS in fungal and metazoan counterparts.

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KW - Nucleocytoplasmic shuttling

KW - Splicing factor1 protein

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DO - 10.1007/s11816-021-00722-0

M3 - Article

AN - SCOPUS:85118830264

SN - 1863-5466

VL - 15

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JO - Plant Biotechnology Reports

JF - Plant Biotechnology Reports

IS - 6

ER -

Identification of a nuclear localization signal mediating the nuclear import of Arabidopsis splicing factor1

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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