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dc.contributor.advisorMa, Din-Pow
dc.contributor.authorHo, Meng-Hsuan
dc.date2009
dc.date.accessioned2020-05-07T20:12:54Z
dc.date.available2020-05-07T20:12:54Z
dc.identifier.urihttps://hdl.handle.net/11668/17462
dc.description.abstractA cotton fiber cDNA, GhRING1, and its corresponding gene have been cloned and characterized. The GhRING1 gene encodes a RING-type ubiquitin ligase (E3) containing 337 amino acids (aa). The GhRING1 protein contains a RING finger motif with conserved cysteine and histine residues at the C-terminus and is classified as a C3H2C3-type RING protein. Blast searches show that GhRING1 has the highest homology to At3g19950 from Arabidopsis. Real time RT-PCR analysis indicates that the GhRING1 gene is highly expressed in cotton fiber in a developmental manner. The transcript level of the GhRING1 gene reaches a maximum in elongating fibers at 15 DPA. In vitro auto-ubiquitination assays using wheat germ extract and a reconstitution system demonstrate that GhRING1 has the ubiquitin E3 ligase activity. A fiber specific lipid transfer protein 4 (FSltp4) is identified as the target substrate of GhRING1 by using a bacterial two-hybrid system. The binding of GhRING1 and FSltp4 is confirmed by using an in vitro pull down assay and a yeast two-hybrid system. The histochemical GUS assay was performed to analyze tissue specificity of the GhRING1 and At3g19950 promoters in transgenic Arabidopsis plants. The GUS assay shows that the promoter of At3g19950 is highly activated in leaves, roots, trichomes and also in anthers and stigma of flowers. In contrast, the GUS expression directed by the promoter of GhRING1 is only located at stipules and anthers and stigma of flowers. The GhRING1 is the first ubiquitin E3 gene isolated and studied from cotton. Based on the expression pattern of GhRING1, FSltp4, and GhUBC E2s and the identification of a fiber-specific target protein, FSltp4, we propose that protein ubiquitination occurs in fiber and the ubiquitin-proteasome pathway regulates fiber development.
dc.publisherMississippi State University
dc.subject.lcshCotton--Genetics.
dc.subject.lcshUbiquitin.
dc.subject.lcshPlant enzymes--Genetics.
dc.subject.lcshCotton--Physiology.
dc.subject.otherE3
dc.subject.otherUbiquitin Ligase
dc.subject.otherUbiquitin
dc.subject.otherFiber
dc.subject.otherTrichome
dc.subject.otherGossypium hirsutum
dc.subject.otherCotton
dc.titleCharacterization and Functional Analysis of a Cotton RING-type Ubiquitin Ligase (E3) Gene
dc.typeDissertation
dc.publisher.departmentDepartment of Biochemistry and Molecular Biology.
dc.date.authorbirth1979
dc.subject.degreeDoctor of Philosophy
dc.subject.majorMolecular Biology
dc.contributor.committeeBoyle, John A.
dc.contributor.committeeJenkins, Johnie
dc.contributor.committeeSaha, Sukumar
dc.contributor.committeePeng, Zhaohua
dc.rights.embargodescriptionMSU Only Indefinitely


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