探讨了JWA在细胞内的分布特征，特别是在有丝分裂过程中的动态变化及与α-微管蛋白的关系。用免疫共沉淀方法研究JWA与α-微管蛋白的相关性；用基因转染技术研究JWA蛋白高表达后JWA蛋白和仅α-微管蛋白的相互关系；用荧光显徽镜技术研究低温处理、药物阻滞细胞周期、JWA反义寡核苷酸处理的PC12细胞JWA蛋白和α-微管蛋白的相互怍用；分别用流式细胞仪分析和激光共聚焦技术检测PC12细胞的各细胞周期蛋白在细胞内的分布。JWA作为一种新的微管相关蛋白，在微管动力学变化过程和细胞周期不同阶段与α-微管蛋白分布平行，可能对微管具有稳定作用。

探讨了JWA蛋白在细胞内的确切定位、与微管蛋白的相互关系及对细胞内氨基酸平衡的调节作用．应用4～37℃（微管解聚-聚合）交替作用法提取纯化大鼠脑组织中微管及其相关蛋白；用免疫共沉淀法研究JWA蛋白与微管蛋白的相互作用；应用基因转染和免疫荧光等实验方法研究低温(4℃)、秋水仙素(1×10-5，5×10-5mol/L)等处理的HBE细胞/NIH3T3细胞中JWA和微管的动态变化及相互关系，应用反义寡核苷酸技术结合氨基酸分析技术探讨JWA蛋白对PC12细胞内氨基酸平衡的调节作用，结果表明，JWA蛋白是一种新的微管相关蛋白，与微管蛋白分布基本平行，在绝大多数情况下伴随微管动力学改变（聚合或解聚）而同步发生变化，并且可能参与了细胞有丝分裂的过程．JWA蛋白是细胞内氨基酸总量的一种重要负性调节蛋白，并选择性地调节细胞内谷氨酸和牛磺酸含量.

Our previous studies demonstrated that JWA, a novel retinoic acids responsive and cytoskeleton related gene, is associated with cell differentiation and apoptosis. In the present study, to elucidate if the JWA is a novel kind of microtubule-associated proteins (MAPs) and functionally link to microtubule, we first successfully identified JWA from the physically purified MAPs complex of rat brain tissues. The results of co-immunoprecipitation, gene transfection and immunofluorescence microscopy assays from HBE and NIH3T3 cells provide strong evidence for a linkage between JWA and β-tubulin. In general, JWA is stably binding to β-tubulin whenever microtubule is polymerized or not, and it may be critical to the mitosis process. In addition, by use of the antisense oligonucleotides technique, we also showed that JWA is a negative modulator on intracellular amino acids in PC12 cells. Further analysis indicated that JWA selectively regulates both taurine, an inhibitory amino acid, and glutamate, an excitatory amino acid. In conclusion, JWA is not only structurally associated, but also a novel functional MAP.

To elucidate the structure characteristic, regulation of expression and the potential function of JWA-a novel retinoic acid responsible and cytoskeleton associate gene, a rat JWA homologue gene and a 621-bp JWA promoter fragment were cloned and analyzed. Using reverse transcription-polymerase chain reaction (RT-PCR), JWA mRNA expression in NIH3T3, K562 and human primary acute promyelocytic leukaemia (APL) cells have been investigated after treatment with all trans retinoic acid (ATRA), N-4-hydroxyphenyl-retinamide (4HPR), arsenic trioxide (As2O3), Phorbol-12-myristate-13-acetate (TPA) and dimethyl sulfoxide (DMSO). The results showed that there is a complete TPA responsive element (TRE) existed in the promoter of JWA at −437 to −443 bp; rat JWA homologue gene showed that there are four nucleotides different from human JWA within coding region. After treatment with TPA, an uneven pattern of JWA transcription existed in different cell lines, suggesting that even TPA induces cell differentiation in different cell lines, and it may be referred to different signal pathways. In ATRA pretreated APL cells, all the above chemicals induce cellular differentiation and down regulate JWA transcription in vitro. This suggests that pretreatment of ATRA on APL cells seems a precondition for turning on JWA involved signal pathway.