PUBLICATION
Cloning and Expression Analysis of a Protein Kinase C Gene, PKCmu, and Its Regulation of the Promoter Region in Zebrafish
- Authors
- Chen, J.Y., Wei, C.C., Chiou, M.J., Su, H.Y., and Kuo, C.M.
- ID
- ZDB-PUB-070629-7
- Date
- 2007
- Source
- DNA and cell biology 26(6): 415-424 (Journal)
- Registered Authors
- Keywords
- none
- MeSH Terms
-
- 5' Untranslated Regions
- Amino Acid Sequence
- Animals
- Base Sequence
- Cloning, Molecular
- DNA Primers/genetics
- DNA, Complementary/genetics
- Gene Expression Regulation, Enzymologic/drug effects
- Genes, Reporter
- Green Fluorescent Proteins/genetics
- Green Fluorescent Proteins/metabolism
- Hormones/pharmacology
- Humans
- Molecular Sequence Data
- Nuclear Localization Signals/chemistry
- Nuclear Localization Signals/genetics
- Promoter Regions, Genetic
- Protein Kinase C/chemistry
- Protein Kinase C/genetics*
- Recombinant Proteins/genetics
- Recombinant Proteins/metabolism
- Sequence Homology, Amino Acid
- Starvation/enzymology
- Starvation/genetics
- Zebrafish/genetics*
- Zebrafish/metabolism
- Zebrafish Proteins/chemistry
- Zebrafish Proteins/genetics*
- PubMed
- 17570765 Full text @ DNA Cell Biol.
Citation
Chen, J.Y., Wei, C.C., Chiou, M.J., Su, H.Y., and Kuo, C.M. (2007) Cloning and Expression Analysis of a Protein Kinase C Gene, PKCmu, and Its Regulation of the Promoter Region in Zebrafish. DNA and cell biology. 26(6):415-424.
Abstract
The cDNA and genomic DNA of zebrafish (Danio rerio) protein kinase Cmu (PKCmu), with its promoter region, were obtained. The 508-amino acid zebrafish PKCmu has 86.17% similarity to human PKCmu. Real-time reverse-transcription polymerase chain reaction analysis with starvation and hormonal treatment found significant differences between the control group and the experimental group after 14 days of starvation. After injecting insulin-like growth factor II (IGF-II), growth hormone (GH), insulin, or human chorionic gonadotropin, significant differences were observed between the control and experimental groups 24 h after treatment. After injecting the gonadotropin-releasing hormone or luteotropin-releasing hormone, significant differences were seen between the control and experimental groups 15 h after treatment. These results suggest that in vivo PKCmu expression is regulated by the insulin family or by the GH, but other sex hormones produced a significant expression level more quickly than the insulin family and GH. The zebrafish PKCmu gene is located on zebrafish chromosome 17 and consists of 16 exons. A 2.6 kilobase pair on the 5' flanking region displayed maximal promoter activity in the zebrafish liver (ZFL) cell line after treatment with IGF-I, IGF-II, and GH. However, a 1.6 kilobase pair on the 5' flanking region displayed maximal promoter activity in the HeLa cell line after treatment with IGF-I, IGF-II, and GH. Finally, PKCmu may have important nuclear effects on cell growth and may involve nuclear localization. By transiently transfecting ZFL cells with various zebrafish PKCmu segments, we identified a nuclear localization signal: the amino acid sequence between amino acids 206 and 209 was able to predominantly direct enhanced green fluorescence protein (EGFP) into the nucleus, whereas a deletion of this motif abrogated the nuclear localization property.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping