微管尖端蛋白在鈣離子感測蛋白基質交互因子活化過程中扮演的角色

劉育伶

doi:10.6844/NCKU.2013.01349

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微管尖端蛋白在鈣離子感測蛋白基質交互因子活化過程中扮演的角色

Microtubule-dependent regulation of ER Ca2+ sensor: the role of microtubule end-binding-protein 1

劉育伶 , Masters　　Advisor：沈孟儒　　

英文 DOI： 10.6844/NCKU.2013.01349

鈣離子； SOCE ； STIM1 ； EB1 ；微管；子宮頸癌； Ca2+ ； SOCE ； STIM1 ； EB1 ； microtubule ； cervical cancer

Abstract │ Reference (88) │ Altmetrics

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Store-operated calcium entry (SOCE) is the major Ca2+ entry mechanism in non-excitable cells and plays an important role in a wide variety of cellular functions, including adhesion, motility, gene expression and proliferation. Stromal interaction molecule 1 (STIM1) is an endoplasmic reticulum (ER) Ca2+ sensor which is essential for SOCE. End-binding-protein 1 (EB1), a microtubule plus-end binding protein (+TIP), controls microtubule dynamics and interacts with most other known +TIPs to target them to growing microtubule plus-ends. Previous studies indicated that microtubules play a facilitative role in SOCE by optimizing the localization of STIM1. However, the mechanisms of microtubule in STIM1 membrane trafficking remain controversial. My study aims to investigate the roles of microtubule-associated proteins on the STIM1-mediated SOCE Ca2+ signal. In order to study the role of EB1 on STIM1 membrane trafficking, I used genetic approach. Results of immunofluorescent images showed that EB1 knockdown altered STIM1 membrane trafficking. Consistently, the results of single cell intracellular Ca2+ measurement showed that EB1 knockdown inhibited the activation of SOCE. Taken these results together, I suggest that EB1 is necessary for STIM1 membrane trafficking. Previous study showed that acetylated--tubulin levels were reduced in EB1-silencing condition. Moreover, histone deacetylase 6 (HDAC6), a member of the class II histone deacetylase that deacetylates microtubule, can alter the microtubule stability and play a role in STIM1 activation. To investigate whether HDAC6 plays a role in EB1-STIM1 interaction, I used tubastatin A, a potent and selective HDAC6 inhibitor, and found that HDAC6 inhibitor reduced the level of juxta-membrane STIM1, but did not affect the STIM1-EB1 interaction. Taken together, I suggest that end-binding protein 1 is necessary for STIM1 trafficking and is involved in the regulation of SOCE activation in cervical cancer cells. Therefore, EB1 may be a potential therapeutic target for tumor malignant behaviors.

Reference ( 88 ) 〈TOP〉

Asahi, M., Sugita, Y., Kurzydlowski, K., De Leon, S., Tada, M., Toyoshima, C., and MacLennan, D. H. (2003) Sarcolipin regulates sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) by binding to transmembrane helices alone or in association with phospholamban. Proceedings of the National Academy of Sciences of the United States of America 100, 5040-5045.
連結：
Akhmanova, A., Steinmetz, M.O. (2010) Microtubule +TIPs at a glance. Journal of Cell Science 123, 3415-3419.
連結：
Aldana-Masangkay, G.I., Sakamoto, K.M. (2011) The role of HDAC6 in cancer. Journal of Biomedicine and Biotechnology 2011.
連結：
Brandl, C.J., Green, N.M., Korczak, B., and MacLennan, D.H. (1986) Two Ca2+ ATPase genes: homologies and mechanistic implications of deduced amino acid sequences. Cell 44, 597-607.
連結：
Berridge, M.J., Lipp, P., Bootman, M.D. (2000) The versatility and universality of calcium signalling. Nature Reviews Molecular Cell Biology 1(1), 11-21.
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