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A Ser/Thr protein kinase phosphorylates MA-ACS1 (Musa acuminata 1-aminocyclopropane-1-carboxylic acid synthase 1) during banana fruit ripening

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Abstract

1-Aminocyclopropane-1-carboxylic acid synthase (ACS) catalyzes the rate-limiting step in ethylene biosynthesis during ripening. ACS isozymes are regulated both transcriptionally and post-translationally. However, in banana, an important climacteric fruit, little is known about post-translational regulation of ACS. Here, we report the post-translational modification of MA-ACS1 (Musa acuminata ACS1), a ripening inducible isozyme in the ACS family, which plays a key role in ethylene biosynthesis during banana fruit ripening. Immunoprecipitation analyses of phospholabeled protein extracts from banana fruit using affinity-purified anti-MA-ACS1 antibody have revealed phosphorylation of MA-ACS1, particularly in ripe fruit tissue. We have identified the induction of a 41-kDa protein kinase activity in pulp at the onset of ripening. The 41-kDa protein kinase has been identified as a putative protein kinase by MALDI–TOF/MS analysis. Biochemical analyses using partially purified protein kinase fraction from banana fruit have identified the protein kinase as a Ser/Thr family of protein kinase and its possible involvement in MA-ACS1 phosphorylation during ripening. In vitro phosphorylation analyses using synthetic peptides and site-directed mutagenized recombinant MA-ACS1 have revealed that serine 476 and 479 residues at the C-terminal region of MA-ACS1 are phosphorylated. Overall, this study provides important novel evidence for in vivo phosphorylation of MA-ACS1 at the molecular level as a possible mechanism of post-translational regulation of this key regulatory protein in ethylene signaling pathway in banana fruit during ripening.

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Abbreviations

ACC:

1-Aminocyclopropane-1-carboxylic acid

ACS:

ACC synthase

CPZ:

Chlorpromazine

DAH:

Days after harvest

DPA:

Days post-anthesis

PAO:

Phenylarsine oxide

PEG:

Polyethylene glycol

TFP:

Trifluoperazine

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Acknowledgments

This work was supported by a research Grant from the Council of Scientific and Industrial Research, Govt. of India to DNS. We thank Mr. Jadav Kumar Ghosh for providing the necessary technical support. The authors are thankful to Dr. Prosenjit Saha CNCI, West Bengal, India for providing the anti-phospho antibodies.

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Author notes

  1. Swarup Roy Choudhury

    Present address: Donald Danforth Plant Science Center, 975 North Warson Road, St. Louis, MO, 63132, USA

  2. Sujit Roy

    Present address: Department of Molecular Genetics, Ohio State University, Columbus, OH, 43210, USA

Authors and Affiliations

  1. Division of Plant Biology, Bose Institute, 93/1, Acharya Prafulla Chandra Road, Kolkata, 700 009, West Bengal, India

    Swarup Roy Choudhury & Dibyendu N. Sengupta

  2. Department of Chemistry, Bose Institute, 93/1, Acharya Prafulla Chandra Road, Kolkata, 700 009, West Bengal, India

    Sujit Roy

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  1. Swarup Roy Choudhury
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  2. Sujit Roy
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  3. Dibyendu N. Sengupta
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Correspondence to Swarup Roy Choudhury.

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S. R. Choudhury and S. Roy have contributed equally to this work.

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Choudhury, S.R., Roy, S. & Sengupta, D.N. A Ser/Thr protein kinase phosphorylates MA-ACS1 (Musa acuminata 1-aminocyclopropane-1-carboxylic acid synthase 1) during banana fruit ripening. Planta 236, 491–511 (2012). https://doi.org/10.1007/s00425-012-1627-9

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  • DOI: https://doi.org/10.1007/s00425-012-1627-9

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