Skip to Main content Skip to Navigation
Theses

Identification et étude du rôle des protéines cibles du monoxyde d'azote (NO) dans les réponses de défense chez le tabac

Abstract : Studies conducted over the past ten years indicate that nitric oxide (NO) is a physiological mediator involved in many physiological processes in plants, including germination, root development, stomatal closure or responses against biotic or abiotic stresses. Despite this important range of functions, the mechanisms underlying the effects of NO in plants remain largely unknown. The present work aims at identifying and functionally characterizing NO target proteins in tobacco in the context of biotic and abiotic stresses. We demonstrated that cryptogein, an elicitor of defense responses, induces a rapid and transient S-nitrosylation of several proteins in tobacco cell suspensions. After purification, a dozen of these proteins have been identified through mass spectrometry analysis. These proteins include CDC48 (Cell Division Cycle 48), a chaperone-like protein belonging to the AAA-ATPase family. The regulation of CDC48 by NO was deeply investigated using a combination of structural and biochemical analyses. Once the in vitro S-nitrosylation of CDC48 was confirmed, we next demonstrated that this process does not affect the secondary structure of the protein but induces local changes in its tertiary structure together with an inhibition of its ATPase activity. The cysteine residue 526, located in the second ATPase domain of the protein, was identified as a probable S-nitrosylation site. This crucial localization may explain the inhibitory effect of NO on CDC48 enzymatic activity. The last part of this work was focused on the analysis of the mechanisms underlying the NO-dependent activation of the protein kinase NtOSAK (Nicotiana tabacum stress activated protein kinase) in tobacco. We demonstrated that NtOSAK forms a constitutive complex with glyceraldehyde 3-phosphate dehydrogenase (GAPDH). In response to salt stress, NO promotes the activation of NtOSAK via the phosphorylation of two serine residues located in the activation loop of the enzyme. Moreover, it induces a rapid S-nitrosylation of GAPDH. Interestingly, this latter process does not affect the complex formation. Our hypothesis is that once S-nitrosylated, GAPDH might act as a phosphorelay recruiting protein substrates for NtOSAK.
Document type :
Theses
Complete list of metadatas

https://tel.archives-ouvertes.fr/tel-00671188
Contributor : Abes Star :  Contact
Submitted on : Thursday, February 16, 2012 - 6:22:27 PM
Last modification on : Saturday, June 6, 2020 - 8:09:01 AM
Long-term archiving on: : Thursday, May 17, 2012 - 2:40:46 AM

File

these_A_ASTIER_Jeremy_2011.pdf
Version validated by the jury (STAR)

Identifiers

  • HAL Id : tel-00671188, version 1
  • PRODINRA : 244440

Collections

Citation

Jérémy Astier. Identification et étude du rôle des protéines cibles du monoxyde d'azote (NO) dans les réponses de défense chez le tabac. Sciences agricoles. Université de Bourgogne, 2011. Français. ⟨NNT : 2011DIJOS015⟩. ⟨tel-00671188⟩

Share

Metrics

Record views

1303

Files downloads

7070