Open Access Research article

Interactions of an Arabidopsis RanBPM homologue with LisH-CTLH domain proteins revealed high conservation of CTLH complexes in eukaryotes

Eva Tomaštíková1, Věra Cenklová2, Lucie Kohoutová3, Beáta Petrovská1, Lenka Váchová2, Petr Halada3, Gabriela Kočárová3 and Pavla Binarová3*

Author Affiliations

1 Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany AS CR, v.v.i., Sokolovská 6, Olomouc, 772 00, Czech Republic

2 Institute of Experimental Botany, AS CR, v.v.i., Sokolovská 6, 772 00, Olomouc, Czech Republic

3 Institute of Microbiology, AS CR, v.v.i., Vídeňská 1083, 142 20, Prague 4, Czech Republic

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BMC Plant Biology 2012, 12:83  doi:10.1186/1471-2229-12-83

Published: 7 June 2012

Additional files

Additional file 1:

Multiple sequence alignment of AtRanBPM plant homologues. Sequence alignment of Arabidopsis RanBPM with Ricinus communisVitis viniferaPopulus trichocarpaSorghum bicolorOryza sativa and Zea mays homologues. Alignment was done using ClustalX2 software [59]. Sequence data of this alignment can be found at accession numbers [Swiss-Prot:F4HYD7] for At1g35470, [Swiss-Prot:B9S762] for R. communis, [Swiss-Prot:F6HWC3] for V. vinifera, [Swiss-Prot:B9MWC1] for P. trichocarpa, [Swiss-Prot:C5XUT1] for S. bicolor, [Swiss-Prot:Q6ZI83] for O. sativa, [Swiss-Prot:B6UAR9] for Z. mays.

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Additional file 2:

Identities and similarities of conserved domains between Arabidopsis RanBPM and its human homologues. Sequence alignment was done for conserved domains SPRY (A), LisH (B), CTLH (C) and CRA (D) of full-sized 90 kDa form (RanBPM_90) and 55 kDa form (RanBPM_55) of human RanBPM protein, and RanBP10 with Arabidopsis RanBPM using ClustalX2 software [59]. Sequence data of this alignment can be found at accession numbers [Swiss-Prot:Q6VN20] for RanBP10, [Swiss-Prot:Q96S59] for 90 kDa RanBPM and [EMBL:BAA23216] for 55 kDa RanBPM. The sequence of the SPRY domain was encountered from the 99 amino acids in AtRanBPM sequence. E- Levels of identities and similarities in amino acid composition of conserved domains between AtRanBPM and its human homologues RanBPM and RanBP10.

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Additional file 3:

Phylogenetic analysis of AtRanBPM and its homologues from other eukaryotic species. The tree was constructed by the neighbor-joining method with the MEGA 5.05 software [23]. Branch numbers represent the percentage of bootstrap values in 1000 sampling replicates. The protein accession numbers are [Swiss-prot:F4HYD7] for AtRanBPM At1g35470, [Swiss-prot: Q9SMS1] At4g09340 (segmental genome duplication of chromosome 1), [Swiss-Prot:B9S762] for R. communis, [Swiss-Prot:F6HWC3] for V. vinifera, [Swiss-Prot:B9MWC1] for P. trichocarpa, [Swiss-Prot:C5XUT1] for S. bicolor, [Swiss-Prot:Q6ZI83] for O. sativa, [Swiss-Prot:B6UAR9] for Z. mays, [Swiss-prot: Q6VN20] for human RanBP10, [Swiss-prot: A3KMV8] for RanBP10 from Bos taurus, [Swiss-prot: B5LX41] for RanBP10 from Felis catus, [Swiss-prot: Q6VN19] for RanBP10 from Mus musculus, [Swiss-prot: Q1LUS8] for RanBP10 from Danio rerio, [Swiss-prot: Q9PTY5] for RanBP9 from Xenopus laevis, [Swiss-prot: Q96S59] for human RanBP9, [Swiss-prot: P69566] for RanBP9 from Mus musculus, [Swiss-prot: Q4Z8K6] for RanBP9/10 from Drosophila melanogaster and [Swiss-prot: P53076] for Gid1/Vid30 homologue from Saccharomyces cerevisiae. Distance bars are given bottom left and bootstrap values are indicated at the nodes.

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Additional file 4:

Immunopurification of GFP-AtRanBPM protein. Immunopurification of GFP-AtRanBPM from extracts of GFP-AtRanBPM expressing cell cultures (IP GFP-RanBPM). GFP immunopurification from extracts of wild type Ler Arabidopsis cells (IP WT) was used as a negative control. A- Proteins were silver stained after separation on SDS-PAGE. Bands corresponding to MW similar of the proteins copurified with GFP-AtRanBPM (IP GFP-RanBPM) were not present in the negative control (IP WT). B- Signal for AtRanBPM was absent in the negative control (IP WT) after detection with anti-AtRanBPM antibody on Western blots. C- Proteins identified by MALDI-MS in negative control (IP WT in A) were background contamination.

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Additional file 5:

Identities and similarities between proteins copurifying with AtRanBPM and human CTLH complex members. Identities and similarities between Arabidopsis and human proteins were analysed in WU-BLAST.

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Additional file 6:

Additional proteins copurified with AtRanBPM. The proteins were identified by LC-MALDI-MS/MS and the identity of the matched peptides was confirmed by high-resolution MALDI-FTMS with mass accuracy below 1 ppm.

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Additional file 7:

GFP-AtRanBPM inArabidopsisroot cells. AtRanBPM-GFP signal is dynamic and moving with the cytoplasmic stream.

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Additional file 8:

Cellular localization of C-terminal GFP and N-terminal GFP AtRanBPM fusion proteins. Cells of Arabidopsis expressing C-terminal GFP AtRanBPM (AtRanBPM-GFP) showed weak cytoplasmic and nuclear signal and accumulation of perinuclear GFP signal similarly as observed for N-terminal GFP AtRanBPM (GFP-AtRanBPM).

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