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Plant Cell Reports

, Volume 36, Issue 7, pp 1113–1123 | Cite as

The Arabidopsis splicing factors, AtU2AF65, AtU2AF35, and AtSF1 shuttle between nuclei and cytoplasms

  • Hyo-Young Park
  • Keh Chien Lee
  • Yun Hee Jang
  • Soon-Kap Kim
  • May Phyo Thu
  • Jeong Hwan LeeEmail author
  • Jeong-Kook KimEmail author
Original Article

Abstract

Key message

The Arabidopsis splicing factors, AtU2AF65, AtU2AF35, and AtSF1 shuttle between nuclei and cytoplasms. These proteins also move rapidly and continuously in the nuclei, and their movements are affected by ATP depletion.

Abstract

The U2AF65 proteins are splicing factors that interact with SF1 and U2AF35 proteins to promote U2snRNP for the recognition of the pre-mRNA 3′ splice site during early spliceosome assembly. We have determined the subcellular localization and movement of these proteins’ Arabidopsis homologs. It was found that Arabidopsis U2AF65 homologs, AtU2AF65a, and AtU2AF65b proteins interact with AtU2AF35a and AtU2AF35b, which are Arabidopsis U2AF35 homologs. We have examined the mobility of these proteins including AtSF1 using fluorescence recovery after photobleaching and fluorescence loss in photobleaching analyses. These proteins displayed dynamic movements in nuclei and their movements were affected by ATP depletion. We have also demonstrated that these proteins shuttle between nuclei and cytoplasms, suggesting that they may also function in cytoplasm. These results indicate that such splicing factors show very similar characteristics to their human counterparts, suggesting evolutionary conservation.

Keywords

Dynamic movement FRAP FLIP SF1 Shuttling U2AF35 U2AF65 

Abbreviations

ATP

Adenosine triphosphate

FLIP

Fluorescence loss in photobleaching

FRAP

Fluorescence recovery after photobleaching

GFP

Green fluorescence protein

LMB

Leptomycin B

NaN3

Sodium azide

Pre-mRNA

Precursor messenger RNA

SF1

Splicing factor 1

snRNP

Small nuclear ribonucleoproteins

U2AFs

U2 snRNP auxiliary splicing factor

NES

Nuclear export signal

Notes

Acknowledgements

This work was supported by the Basic Science Research Program through the National Research Foundation of Korea, funded by the Ministry of Education (to J.-K. Kim), and by a Korea University Grant (to J.-K. Kim).

Compliance with ethical standards

Conflict of interest

The authors have no conflicts of interest to declare.

Supplementary material

299_2017_2142_MOESM1_ESM.avi (24.3 mb)
Figure S1. Time-lapse movies derived from the FLIP experiment shown in Fig. 5.Table S1. Primers used in this study. (AVI 24841 kb)
299_2017_2142_MOESM2_ESM.avi (33.2 mb)
Supplementary material 2 (AVI 33984 kb)
299_2017_2142_MOESM3_ESM.docx (14 kb)
Supplementary material 3 (DOCX 14 kb)
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Supplementary material 4 (AVI 26524 kb)
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Supplementary material 5 (AVI 24516 kb)
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Supplementary material 6 (AVI 26298 kb)
299_2017_2142_MOESM7_ESM.avi (25.4 mb)
Supplementary material 7 (AVI 26043 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Hyo-Young Park
    • 1
  • Keh Chien Lee
    • 1
  • Yun Hee Jang
    • 1
  • Soon-Kap Kim
    • 2
  • May Phyo Thu
    • 1
  • Jeong Hwan Lee
    • 3
    Email author
  • Jeong-Kook Kim
    • 1
    Email author
  1. 1.Department of Life SciencesKorea UniversitySeoulRepublic of Korea
  2. 2.Division of Biological and Environmental Sciences and Engineering, Center for Desert AgricultureKing Abdullah University of Science and TechnologyThuwalSaudi Arabia
  3. 3.Department of Life SciencesChonbuk National UniversityJeonju-SiRepublic of Korea

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