Characterization and promoter analysis of a cotton RING-type ubiquitin ligase (E3) gene.
A cotton fiber cDNA, GhRING1, and its corresponding gene have been cloned and characterized. The GhRING1 gene encodes a RING-type ubiquitin ligase (E3) containing 338 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 C(3)H(2)C(3)-type RING protein. Blast searches show that GhRING1 has the highest homology to At3g19950, a zinc finger family protein from Arabidopsis. Real time RT-PCR analysis indicates that the GhRING1 gene is expressed in cotton fibers in a developmental manner. The transcript level of GhRING1 gene reaches a maximum in elongating fibers at 15 days post-anthesis (DPA). In vitro auto-ubiquitination assays using wheat germ extract and a reconstitution system demonstrate that GhRING1 has the ubiquitin E3 ligase activity. 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 GhRING1 promoter is only located at stipules and anthers. The expression pattern of GhRING1 suggests that protein ubiquitination and turnover may be involved in transition to different stages of cotton fiber development.
Ho
Meng-Hsuan
MH
Department of Biochemistry and Molecular Biology, Mississippi State University, Mississippi State, Box 9650, MS, 39762, USA. mh277@msstate.edu
Saha
Sukumar
S
Jenkins
Johnie N
JN
Ma
Din-Pow
DP
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics; D003001:Cloning, Molecular; D032561:Cotton Fiber; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D003368:Gossypium / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D044767:Ubiquitin-Protein Ligases / Q000235:genetics*; D054875:Ubiquitination
D044767:Ubiquitin-Protein Ligases