N-Acylethanolamine Metabolism During Seed Germination: Molecular Identification of a Functional N-Acylethanolamine Amidohydrolase Metadata

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Title

  • Main Title N-Acylethanolamine Metabolism During Seed Germination: Molecular Identification of a Functional N-Acylethanolamine Amidohydrolase

Creator

  • Author: Shrestha, Rhidaya
    Creator Type: Personal

Contributor

  • Chair: Chapman, Kent D.
    Contributor Type: Personal
    Contributor Info: Major Professor
  • Committee Member: Pirtle, Robert M.
    Contributor Type: Personal
  • Committee Member: Knesek, John
    Contributor Type: Personal
  • Committee Member: Dickstein, Rebecca
    Contributor Type: Personal
  • Committee Member: Root, Douglas D.
    Contributor Type: Personal

Publisher

  • Name: University of North Texas
    Place of Publication: Denton, Texas

Date

  • Creation: 2004-08
  • Digitized: 2007-12-04

Language

  • English

Description

  • Content Description: N-Acylethanolamines (NAEs) are endogenous lipid metabolites that occur in a variety of dry seeds, and their levels decline rapidly during the first few hours of imbibition (Chapman et al., 1999, Plant Physiol., 120:1157-1164). Biochemical studies supported the existence of an NAE amidohydrolase activity in seeds and seedlings, and efforts were directed toward identification of DNA sequences encoding this enzyme. Mammalian tissues metabolize NAEs via an amidase enzyme designated fatty acid amide hydrolase (FAAH). Based on the characteristic amidase signature sequence in mammalian FAAH, a candidate Arabidopsis cDNA was identified and isolated by reverse transcriptase-PCR. The Arabidopsis cDNA was expressed in E. coli and the recombinant protein indeed hydrolyzed a range of NAEs to free fatty acids and ethanolamine. Kinetic parameters for the recombinant protein were consistent with those properties of the rat FAAH, supporting identification of this Arabidopsis cDNA as a FAAH homologue. Two T-DNA insertional mutant lines with disruptions in the Arabidopsis NAE amidohydrolase gene (At5g64440) were identified. The homozygous mutant seedlings were more sensitive than the wild type to exogenously applied NAE 12:0. Transgenic seedlings overexpressing the NAE amidohydrolase enzyme showed noticeably greater tolerance to NAE 12:0 than wild type seedlings. These results together provide evidence in vitro and in vivo for the molecular identification of Arabidopsis NAE amidohydrolase. Moreover, the plants with altered NAE amidohydrolase expression may provide new tools for improved understanding of the role of NAEs in germination and seedling growth.

Subject

  • Library of Congress Subject Headings: Amidases.
  • Library of Congress Subject Headings: Germination.
  • Keyword: N-acylethanolamine
  • Keyword: NAE metabolism
  • Keyword: NAE amidohydrolase
  • Keyword: seed germination
  • Keyword: AtFAAH

Collection

  • Name: UNT Theses and Dissertations
    Code: UNTETD

Institution

  • Name: UNT Libraries
    Code: UNT

Rights

  • Rights Access: public
  • Rights License: copyright
  • Rights Holder: Shrestha, Rhidaya
  • Rights Statement: Copyright is held by the author, unless otherwise noted. All rights reserved.

Resource Type

  • Thesis or Dissertation

Format

  • Text

Identifier

  • OCLC: 56821055
  • Archival Resource Key: ark:/67531/metadc4575

Degree

  • Degree Name: Doctor of Philosophy
  • Degree Level: Doctoral
  • Degree Discipline: Biochemistry
  • Academic Department: Department of Biological Sciences
  • Degree Grantor: University of North Texas

Note