Genetic disruption of ATAT1 causes RhoA downregulation through abnormal truncation of C/EBPβ
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Microtubule acetylation has been shown to regulate actin filamentdynamics by modulating signaling pathways that controlactin organization, although the precise mechanisms remainunknown. In this study, we found that the downregulation ofmicrotubule acetylation via the disruption ATAT1 (which encodesα-tubulin N-acetyltransferase 1) inhibited the expressionof RhoA, a small GTPase involved in regulating the organizationof actin filaments and the formation of stress fibers. Analysisof RHOA promoter and chromatin immunoprecipitationassays revealed that C/EBPβ is a major regulator of RHOA expression. Interestingly, the majority of C/EBPβ in ATAT1 knockout(KO) cells was found in the nucleus as a 27-kDa fragment(referred to as C/EBPβp27) lacking the N-terminus of C/EBPβ. Overexpression of a gene encoding a C/EBPβp27-mimicking proteinvia an N-terminal deletion in C/EBPβ led to competitivebinding with wild-type C/EBPβ at the C/EBPβ binding site inthe RHOA promoter, resulting in a significant decrease of RHOAexpression. We also found that cathepsin L (CTSL), which isoverexpressed in ATAT1 KO cells, is responsible for C/EBPβp27formation in the nucleus. Treatment with a CTSL inhibitor ledto the restoration of RHOA expression by downregulation ofC/EBPβp27 and the invasive ability of ATAT1 KO MDA-MB-231breast cancer cells. Collectively, our findings suggest that thedownregulation of microtubule acetylation associated with ATAT1deficiency suppresses RHOA expression by forming C/EBPβp27in the nucleus through CTSL. We propose that CTSL and C/EBPβp27may represent a novel therapeutic target for breast cancertreatment.
Abstract
Microtubule acetylation has been shown to regulate actin filamentdynamics by modulating signaling pathways that controlactin organization, although the precise mechanisms remainunknown. In this study, we found that the downregulation ofmicrotubule acetylation via the disruption ATAT1 (which encodesα-tubulin N-acetyltransferase 1) inhibited the expressionof RhoA, a small GTPase involved in regulating the organizationof actin filaments and the formation of stress fibers. Analysisof RHOA promoter and chromatin immunoprecipitationassays revealed that C/EBPβ is a major regulator of RHOA expression. Interestingly, the majority of C/EBPβ in ATAT1 knockout(KO) cells was found in the nucleus as a 27-kDa fragment(referred to as C/EBPβp27) lacking the N-terminus of C/EBPβ. Overexpression of a gene encoding a C/EBPβp27-mimicking proteinvia an N-terminal deletion in C/EBPβ led to competitivebinding with wild-type C/EBPβ at the C/EBPβ binding site inthe RHOA promoter, resulting in a significant decrease of RHOAexpression. We also found that cathepsin L (CTSL), which isoverexpressed in ATAT1 KO cells, is responsible for C/EBPβp27formation in the nucleus. Treatment with a CTSL inhibitor ledto the restoration of RHOA expression by downregulation ofC/EBPβp27 and the invasive ability of ATAT1 KO MDA-MB-231breast cancer cells. Collectively, our findings suggest that thedownregulation of microtubule acetylation associated with ATAT1deficiency suppresses RHOA expression by forming C/EBPβp27in the nucleus through CTSL. We propose that CTSL and C/EBPβp27may represent a novel therapeutic target for breast cancertreatment.
참고문헌 (19)
- Humphrey JD, "Mechanotransduction and extracellular matrix homeostasis", Nat Rev Mol Cell Biol, 15, pp.802~812 (2014)
- Seetharaman S, "Cytoskeletal crosstalk in cell migration", Trends Cell Biol, 30, pp.720~735 (2020)
- Garcin C, "Microtubules in cell migration", Essays Biochem, 63, pp.509~520 (2019)
- Inoue D, "Actin filaments regulate microtubule growth at the centrosome", EMBO J, 38, pp.e99630~ (2019)
- Dubey T, "Photodynamic sensitizers modulate cytoskeleton structural dynamics in neuronal cells", Cytoskeleton(Hoboken), 78, pp.232~248 (2021)
- Fife CM, "Movers and shakers : cell cytoskeleton in cancer metastasis", Br J Pharmacol, 171, pp.5507~5523 (2014)
- O’Connor K, "Dynamic functions of RhoA in tumor cell migration and invasion", Small GTPases, 4, pp.141~147 (2013)
- Chan CH, "Deciphering the transcriptional complex critical for RhoA gene expression and cancer metastasis", Nat Cell Biol, 12, pp.457~467 (2010)
- Nomikou E, "Functional analysis of the promoters of the small GTPases RhoA and RhoB in embryonic stem cells", Biochem Biophys Res Commun, 491, pp.754~759 (2017)
- Bundy LM, "CCAAT", enhancer binding protein beta(C, Oncogene, pp.2003, 22[DOI]
- Oh S, "Genetic disruption of tubulin acetyltransferase, alphaTAT1, inhibits proliferation and invasion of colon cancer cells through decreases in Wnt1", beta-catenin signaling, 2017, pp.482, 8~14
- Ko P, "Microtubule acetylation controls MDA-MB-231 breast cancer cell invasion through the modulation of endoplasmic reticulum stress", Int J Mol Sci, 22, pp.6018~ (2021)
- Zahnow CA, "CCAAT", enhancer-binding protein beta : its role in breast cancer and associations with receptor tyrosine kinases, 2009, pp.11, e12~
- Park JS, "The effect of matrix stiffness on the differentiation of mesenchymal stem cells in response to TGF-beta", Biomaterials, 32, pp.3921~3930 (2011)
- Williams SC, "CRP2(C", EBP beta)contains a bipartite regulatory domain that controls transcriptional activation, DNA binding and cell specificity, 1995, pp.14, 3170~3183
- Sudhan DR, "Cathepsin L targeting in cancer treatment", Pharmacol Ther, 155, pp.105~116 (2015)
- Zhang W, "Overexpression of cysteine cathepsin L is a marker of invasion and metastasis in ovarian cancer", Oncol Rep, 31, pp.1334~1342 (2014)
- Goulet B, "A novel proteolytically processed CDP", Cux isoform of 90 kDa is generated by cathepsin L, 2006, pp.387, 1285~1293
- Reed NA, "Microtubule acetylation promotes kinesin-1 binding and transport", Curr Biol, 16, pp.2166~2172 (2006)