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Predominantly Cytoplasmic Localization in Yeast of ASR1, a Non-Receptor Transcription Factor from Plants.

Urtasun N, Correa García S, Iusem ND, Bermúdez Moretti M - Open Biochem J (2010)

Bottom Line: It is associated with water-deficit stress and is involved in adaptation to dry climates.For that purpose, we employed an in vivo eukaryotic expression system, the heterologous model Saccharomyces cerevisiae, including wild type strains as well as mutants in which the variant ASR1 previously proved to be functionally protective against osmotic stress.The results are discussed in terms of a plausible dual (cytoplasmic and nuclear) role of ASR proteins.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Ciudad Universitaria (1428), Buenos Aires, Argentina.

ABSTRACT
The Asr gene family (named after abscisic acid, stress and ripening), currently classified as a novel group of the LEA superfamily, is exclusively present in the genomes of seed plants, except for the Brassicaceae family. It is associated with water-deficit stress and is involved in adaptation to dry climates. Motivated by separate reports depicting ASR proteins as either transcription factors or chaperones, we decided to determine the intracellular localization of ASR proteins. For that purpose, we employed an in vivo eukaryotic expression system, the heterologous model Saccharomyces cerevisiae, including wild type strains as well as mutants in which the variant ASR1 previously proved to be functionally protective against osmotic stress. Our methodology involved immunofluorescence-based confocal microscopy, without artificially altering the native structure of the protein under study. Results show that, in both normal and osmotic stress conditions, recombinant ASR1 turned out to localize mainly to the cytoplasm, irrespective of the genotype used, revealing a scattered distribution in the form of dots or granules. The results are discussed in terms of a plausible dual (cytoplasmic and nuclear) role of ASR proteins.

No MeSH data available.


Amino acid sequence of ASR1, the ASR variant chosen for this study. Highlighted are key residues within the NLS (positions 92-105) [16].
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Figure 1: Amino acid sequence of ASR1, the ASR variant chosen for this study. Highlighted are key residues within the NLS (positions 92-105) [16].

Mentions: The Lycoperson esculentum Asr1 full-length cDNA, containing the NLS (amino acid positions 92-105) (Fig. 1), was subcloned into the Xba1/Kpn1 site of the high-copy-number shuttle vector pYES2 (Invitrogen, CH Groningen, The Netherlands) that contains the GAL1 promoter [18]. Wild type and mutant cells were transformed with the recombinant vector (pYES2-Asr1) using the method described by Chen et al. (1992)[19]. As a control, the same strains were transformed with the empty vector.


Predominantly Cytoplasmic Localization in Yeast of ASR1, a Non-Receptor Transcription Factor from Plants.

Urtasun N, Correa García S, Iusem ND, Bermúdez Moretti M - Open Biochem J (2010)

Amino acid sequence of ASR1, the ASR variant chosen for this study. Highlighted are key residues within the NLS (positions 92-105) [16].
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2908927&req=5

Figure 1: Amino acid sequence of ASR1, the ASR variant chosen for this study. Highlighted are key residues within the NLS (positions 92-105) [16].
Mentions: The Lycoperson esculentum Asr1 full-length cDNA, containing the NLS (amino acid positions 92-105) (Fig. 1), was subcloned into the Xba1/Kpn1 site of the high-copy-number shuttle vector pYES2 (Invitrogen, CH Groningen, The Netherlands) that contains the GAL1 promoter [18]. Wild type and mutant cells were transformed with the recombinant vector (pYES2-Asr1) using the method described by Chen et al. (1992)[19]. As a control, the same strains were transformed with the empty vector.

Bottom Line: It is associated with water-deficit stress and is involved in adaptation to dry climates.For that purpose, we employed an in vivo eukaryotic expression system, the heterologous model Saccharomyces cerevisiae, including wild type strains as well as mutants in which the variant ASR1 previously proved to be functionally protective against osmotic stress.The results are discussed in terms of a plausible dual (cytoplasmic and nuclear) role of ASR proteins.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Ciudad Universitaria (1428), Buenos Aires, Argentina.

ABSTRACT
The Asr gene family (named after abscisic acid, stress and ripening), currently classified as a novel group of the LEA superfamily, is exclusively present in the genomes of seed plants, except for the Brassicaceae family. It is associated with water-deficit stress and is involved in adaptation to dry climates. Motivated by separate reports depicting ASR proteins as either transcription factors or chaperones, we decided to determine the intracellular localization of ASR proteins. For that purpose, we employed an in vivo eukaryotic expression system, the heterologous model Saccharomyces cerevisiae, including wild type strains as well as mutants in which the variant ASR1 previously proved to be functionally protective against osmotic stress. Our methodology involved immunofluorescence-based confocal microscopy, without artificially altering the native structure of the protein under study. Results show that, in both normal and osmotic stress conditions, recombinant ASR1 turned out to localize mainly to the cytoplasm, irrespective of the genotype used, revealing a scattered distribution in the form of dots or granules. The results are discussed in terms of a plausible dual (cytoplasmic and nuclear) role of ASR proteins.

No MeSH data available.