Quantitative proteomics identifies oxidant-induced, AtMPK6-dependent changes in <em>Arabidopsis thaliana</em> protein profiles

Godfrey Miles

Marcus Samuel

Jeffrey Ranish

Sam Donohoe

Gina Sperrazzo

Brian Ellis

Supplementary Material:

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^{Michael Smith Laboratories; University of British Columbia; Vancouver, BC CA}

^{Department of Biological Sciences; University of Calgary; Calgary, AB CA}

^{Institute for Systems Biology; Seattle, WA USA}

^{Corresponding author.}

Correspondence to: Godfrey P. Miles; U.S. Department of Agriculture - Agricultural Research Service; Yakima Agricultural Research Laboratory; 5230 Konnowac Pass Road; Wapato, WA 98951 USA; Tel.: 509.454.4476; Email: /vog.adsu.sra@selim.yerfdog Brian E. Ellis; University of British Columbia; Michael Smith Laboratories; 2185 East Mall; Vancouver, BC V6T 1Z4 CA; Tel.: 604.822.3451; Email: ac.cbu.egnahcretni@eeb

^{Current address: U.S. Department of Agriculture; Agricultural Research Service; Wapato, WA USA}

^{Current address: University of Washington School of Medicine; Seattle, WA USA}

^{Current address: Amgen Incorporated; Department of Pharmaceutics; One Amgen Center Drive; Thousand Oaks, CA USA}

Received 2008 Oct 24; Accepted 2009 Mar 17.

Abstract

In Arabidopsis thaliana, oxidant-induced signalling has been shown to utilize the mitogen-activated protein kinase (MAPK), AtMPK6. To identify proteins whose accumulation is altered by ozone in an AtMPK6-dependent manner we employed isotope-coded affinity tagging (ICAT) technology to investigate the impact of AtMPK6-suppression on the protein profiles in Arabidopsis both before (air control) and during continuous ozone (O₃) fumigation (500 nL L^{for 8 h). Among the 150 proteins positively identified and quantified in the O}₃-treated plants, we identified thirteen proteins whose abundance was greater in the AtMPK6-suppressed genotype than in wild-type (WT). These include the antioxidant proteins, monodehydroascorbate reductase, peroxiredoxin Q, and glutathione reductase. A further eighteen proteins were identified whose abundance was lower in the ozone-treated AtMPK6-suppressed line relative to ozone-exposed WT plants. These predominantly comprised proteins involved in carbohydrate-, energy-, and amino acid metabolism, and tetrapyrrole biosynthesis. In control plants, five proteins increased, and nine proteins decreased in abundance in the AtMPK6-suppressed genotype compared to that of the WT, reflecting changes in the protein composition of plants that have AtMPK6 constitutively suppressed. Since a number of these proteins are part of the redox response pathway, and loss of AtMPK6 renders Arabidopsis more susceptible to oxidative stress, we propose that AtMPK6 plays a key role in the plant's overall ability to manage oxidative stress.

Key words: Arabidopsis thaliana, AtMPK6, isotope-coded affinity tag (ICAT), ozone, MAPK, signalling

Abstract

Acknowledgements

We are thankful to the Institute of Systems Biology (ISB), especially to Professor Ruedi Aebersold, who kindly extended an invitation to work at the ISB making this project possible, and to Jimmy Eng for his invaluable assistance with statistical analysis. This work was supported by the Natural Sciences and Engineering Research Council of Canada.

Acknowledgements

Footnotes

Previously published online as a Plant Signaling & Behavior E-publication: http://www.landesbioscience.com/journals/psb/article/8538

Footnotes

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