A Novel Environmental Responsive Gene JWA Structures And Related Functions

The primary goal of toxicological investigations is to assess the effects of environmental exposure on the health status of an exposed animal or human. Environmental toxicogenomics is a new approach to environmental toxicology. It allows us to identify and characterize genomic signatures of environmental toxicants as gene and protein expression profiles. A lot of human diseases are recognized related to various environmental exposures. A series of environmental responsive genes in cells will actively regulated their expression levels to adapt the changed environmental factors when human exposed to physical, chemical and biological harmful agents. The consequence of environmental exposure is dependent on the exposure levels and the gene-environmental interactions. The genes involved in environmental responsiveness usually refer to those related to cell metabolism, cell differentiation, apoptosis, cell cycle, DNA injury and repairs as well. Using the differential display high-throughput technique, we previously cloned a novel gene JWA (NCBI, AF070523.1) from all-trans retinoid acid (ATRA) induced cell differentiation model. Our primary data showed that JWA is broadly distributed in most of examined eukaryotic cells and tissues and with higher transcription and translation levels. The intracellular distribution of the protein is very similar to that of cytoskeleton under immunofluorescent microscopy. The expressions of JWA in cells are showed actively responsive to the cell differentiation inducers, such as ATRA, TPA (12-O-tetradecanoyl-phorbol-13 acetate), 4HPR (N-4-hydroxyphenyl retinamide) and As2O3 (arsenic trioxide) and the oxidative and heat shock stresses. In this study, we use gene-specific primers and antibody to further investigate the genomic structures, protein structures and the relationships of JWA with cytoskeleton microtubules. At the same time, we pay more attention to explore JWA may referred molecular mechanisms and signal pathways in regulating cell differentiations, apoptosis, and as an environmental responsive gene as well. The techniques or methods used in this study are include molecular cloning, reverse transcriptase-polymerase chain reaction (RT-PCR), gene transfection and knock-down, flowcytometry (FCM), and immuno- fluorescent microscopy (IFM), immuno-precippitation (IP), immunoelectro-microscopy (IEM), immuno blotting and so on.1, For gene structures: use molecular cloning techniques, cell biology experimental techniques and web tools, we identified the genomic, cDNA, and protein structures of JWA; and some JWA homologs either in cDNA or proteins are also found; finally and most importantly, we examined with accumulated evidences that JWA protein is structurally and functionally existed as a novel microtubule associate protein (MAP). These results are described briefly as follows:(1) JWA is located on Chromosome 3 (3p14, NT₀22423.12) in human beings. Several cis-elements are found from its promoter region of within minus 3,000bp such as for stresses, TPA, heat shock, and hemin.(2) The finally confirmed full length cDNA of JWA in human contains 2088bp (the coding region at 90th-65 6th bp) and with a composed of base pairs ratio at 69/31 for at/eg; eleven single nucleotide polymorphism sites (SNP) are identified in its cDNA and two of them are located in JWA coding region (428C/A, 546G/A). Two conserved cDNA domains prenylated rab acceptor (PRA1) and endomembrane protein (EM70) are also found.(3) Within the coding region of 188 amino acids, three transmembrane domains (TMD) are identified and each with 23 amino acids (36-58, 64-86, 108-130). The protein kinase C (PKC) phosphorylational sites are found at the both side of TMD (SDR_SLR). In addition, the secondary structures such as alpha helices, beta-turns, extend strands and random coils are also predicted from JWA protein sequences.(4) Compared to the predicted 21.5 kDa of JWA molecule, the anti- JWA protein C-terminal 20 amino acid peptide serum identified molecule is little bit larger (22.35 kDa). We previ...