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Arabidopsis thaliana AUCSIA-1 regulates auxin biology and physically interacts with a kinesin-related protein.

Molesini B, Pandolfini T, Pii Y, Korte A, Spena A - PLoS ONE (2012)

Bottom Line: Two allelic mutants for AtAucsia-1 gene did not display visible root morphological alterations; however both basipetal and acropetal indole-3-acetic acid (IAA) root transport was reduced as compared with wild-type plants.The transcript steady state levels of the auxin efflux transporters ATP BINDING CASSETTE subfamily B (ABCB) ABCB1, ABCB4 and ABCB19 were reduced in ataucsia-1 plants.Gene ontology predictions for the two proteins are consistent with the hypothesis that the AtAUCSIA-1/AtAUCSIA-1IP complex is involved in the regulation of the cytoskeleton dynamics underlying auxin biology.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology, University of Verona, Verona, Italy.

ABSTRACT
Aucsia is a green plant gene family encoding 44-54 amino acids long miniproteins. The sequenced genomes of most land plants contain two Aucsia genes. RNA interference of both tomato (Solanum lycopersicum) Aucsia genes (SlAucsia-1 and SlAucsia-2) altered auxin sensitivity, auxin transport and distribution; it caused parthenocarpic development of the fruit and other auxin-related morphological changes. Here we present data showing that the Aucsia-1 gene of Arabidopsis thaliana alters, by itself, root auxin biology and that the AtAUCSIA-1 miniprotein physically interacts with a kinesin-related protein. The AtAucsia-1 gene is ubiquitously expressed, although its expression is higher in roots and inflorescences in comparison to stems and leaves. Two allelic mutants for AtAucsia-1 gene did not display visible root morphological alterations; however both basipetal and acropetal indole-3-acetic acid (IAA) root transport was reduced as compared with wild-type plants. The transcript steady state levels of the auxin efflux transporters ATP BINDING CASSETTE subfamily B (ABCB) ABCB1, ABCB4 and ABCB19 were reduced in ataucsia-1 plants. In ataucsia-1 mutant, lateral root growth showed an altered response to i) exogenous auxin, ii) an inhibitor of polar auxin transport and iii) ethylene. Overexpression of AtAucsia-1 inhibited primary root growth. In vitro and in vivo protein-protein interaction experiments showed that AtAUCSIA-1 interacts with a 185 amino acids long fragment belonging to a 2712 amino acids long protein of unknown function (At4g31570). Bioinformatics analysis indicates that the AtAUCSIA-1 interacting protein (AtAUCSIA-1IP) clusters with a group of CENP-E kinesin-related proteins. Gene ontology predictions for the two proteins are consistent with the hypothesis that the AtAUCSIA-1/AtAUCSIA-1IP complex is involved in the regulation of the cytoskeleton dynamics underlying auxin biology.

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Arabidopsis thaliana Aucsia-1 coding sequence and expression in different organs.(A) Pairwise alignments of Aucsia-1 coding sequences from A. thaliana and Solanum Lycopersicon (upper) and comparison of the corresponding amino acid sequences (lower). * identical in all sequences, : for conserved substitutions; . for semi-conserved substitutions. (B) Expression pattern analysis of AtAucsia-1 in various tissues of wild-type adult plants assessed by quantitative real-time PCR (qRT-PCR). The expression levels were normalized using actin as endogenous control gene and the relative expression ratios were calculated using stems as calibrator sample. The values reported are means ± standard error (SE) (n = 3).
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pone-0041327-g001: Arabidopsis thaliana Aucsia-1 coding sequence and expression in different organs.(A) Pairwise alignments of Aucsia-1 coding sequences from A. thaliana and Solanum Lycopersicon (upper) and comparison of the corresponding amino acid sequences (lower). * identical in all sequences, : for conserved substitutions; . for semi-conserved substitutions. (B) Expression pattern analysis of AtAucsia-1 in various tissues of wild-type adult plants assessed by quantitative real-time PCR (qRT-PCR). The expression levels were normalized using actin as endogenous control gene and the relative expression ratios were calculated using stems as calibrator sample. The values reported are means ± standard error (SE) (n = 3).

Mentions: A search for tomato Aucsia-1 homologous gene in the Arabidopsis thaliana genome identified a candidate gene of unknown function (Figure 1A), named AtAucsia-1 (At3g01130), located in the chromosome 3, and annotated as component of the endomembrane system (http://www.arabidopsis.org/). AtAucsia-1 gene comprises 3 exons encoding a peptide of 53 amino acids (5546.5 Da). Quantitative RT-PCR (qRT-PCR), carried out on adult A. thaliana plants, showed that AtAucsia-1 gene is ubiquitously expressed, and yet relatively more abundant in inflorescences and roots (Figure 1B).


Arabidopsis thaliana AUCSIA-1 regulates auxin biology and physically interacts with a kinesin-related protein.

Molesini B, Pandolfini T, Pii Y, Korte A, Spena A - PLoS ONE (2012)

Arabidopsis thaliana Aucsia-1 coding sequence and expression in different organs.(A) Pairwise alignments of Aucsia-1 coding sequences from A. thaliana and Solanum Lycopersicon (upper) and comparison of the corresponding amino acid sequences (lower). * identical in all sequences, : for conserved substitutions; . for semi-conserved substitutions. (B) Expression pattern analysis of AtAucsia-1 in various tissues of wild-type adult plants assessed by quantitative real-time PCR (qRT-PCR). The expression levels were normalized using actin as endogenous control gene and the relative expression ratios were calculated using stems as calibrator sample. The values reported are means ± standard error (SE) (n = 3).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3401106&req=5

pone-0041327-g001: Arabidopsis thaliana Aucsia-1 coding sequence and expression in different organs.(A) Pairwise alignments of Aucsia-1 coding sequences from A. thaliana and Solanum Lycopersicon (upper) and comparison of the corresponding amino acid sequences (lower). * identical in all sequences, : for conserved substitutions; . for semi-conserved substitutions. (B) Expression pattern analysis of AtAucsia-1 in various tissues of wild-type adult plants assessed by quantitative real-time PCR (qRT-PCR). The expression levels were normalized using actin as endogenous control gene and the relative expression ratios were calculated using stems as calibrator sample. The values reported are means ± standard error (SE) (n = 3).
Mentions: A search for tomato Aucsia-1 homologous gene in the Arabidopsis thaliana genome identified a candidate gene of unknown function (Figure 1A), named AtAucsia-1 (At3g01130), located in the chromosome 3, and annotated as component of the endomembrane system (http://www.arabidopsis.org/). AtAucsia-1 gene comprises 3 exons encoding a peptide of 53 amino acids (5546.5 Da). Quantitative RT-PCR (qRT-PCR), carried out on adult A. thaliana plants, showed that AtAucsia-1 gene is ubiquitously expressed, and yet relatively more abundant in inflorescences and roots (Figure 1B).

Bottom Line: Two allelic mutants for AtAucsia-1 gene did not display visible root morphological alterations; however both basipetal and acropetal indole-3-acetic acid (IAA) root transport was reduced as compared with wild-type plants.The transcript steady state levels of the auxin efflux transporters ATP BINDING CASSETTE subfamily B (ABCB) ABCB1, ABCB4 and ABCB19 were reduced in ataucsia-1 plants.Gene ontology predictions for the two proteins are consistent with the hypothesis that the AtAUCSIA-1/AtAUCSIA-1IP complex is involved in the regulation of the cytoskeleton dynamics underlying auxin biology.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology, University of Verona, Verona, Italy.

ABSTRACT
Aucsia is a green plant gene family encoding 44-54 amino acids long miniproteins. The sequenced genomes of most land plants contain two Aucsia genes. RNA interference of both tomato (Solanum lycopersicum) Aucsia genes (SlAucsia-1 and SlAucsia-2) altered auxin sensitivity, auxin transport and distribution; it caused parthenocarpic development of the fruit and other auxin-related morphological changes. Here we present data showing that the Aucsia-1 gene of Arabidopsis thaliana alters, by itself, root auxin biology and that the AtAUCSIA-1 miniprotein physically interacts with a kinesin-related protein. The AtAucsia-1 gene is ubiquitously expressed, although its expression is higher in roots and inflorescences in comparison to stems and leaves. Two allelic mutants for AtAucsia-1 gene did not display visible root morphological alterations; however both basipetal and acropetal indole-3-acetic acid (IAA) root transport was reduced as compared with wild-type plants. The transcript steady state levels of the auxin efflux transporters ATP BINDING CASSETTE subfamily B (ABCB) ABCB1, ABCB4 and ABCB19 were reduced in ataucsia-1 plants. In ataucsia-1 mutant, lateral root growth showed an altered response to i) exogenous auxin, ii) an inhibitor of polar auxin transport and iii) ethylene. Overexpression of AtAucsia-1 inhibited primary root growth. In vitro and in vivo protein-protein interaction experiments showed that AtAUCSIA-1 interacts with a 185 amino acids long fragment belonging to a 2712 amino acids long protein of unknown function (At4g31570). Bioinformatics analysis indicates that the AtAUCSIA-1 interacting protein (AtAUCSIA-1IP) clusters with a group of CENP-E kinesin-related proteins. Gene ontology predictions for the two proteins are consistent with the hypothesis that the AtAUCSIA-1/AtAUCSIA-1IP complex is involved in the regulation of the cytoskeleton dynamics underlying auxin biology.

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