Abstract
Background
Golgi apparatus (GA) is assembled as a crescent-like ribbon in mammalian cells under immunofluorescence microscope without knowing the sha** mechanisms. It is estimated that roughly 1/5 of the genes encoding kinases or phosphatases in human genome participate in the assembly of Golgi ribbon, reflecting protein modifications play major roles in building Golgi ribbon.
Methods
To explore how Golgi ribbon is shaped as a crescent-like structure under the guidance of protein modifications, we identified a protein complex containing the scaffold proteins Ajuba, two known GA regulators including the protein kinase Aurora-A and the protein arginine methyltransferase PRMT5, and the common substrate of Aurora-A and PRMT5, HURP. Mutual modifications and activation of PRMT5 and Aurora-A in the complex leads to methylation and in turn phosphorylation of HURP, thereby producing HURP p725. The HURP p725 localizes to GA vicinity and its distribution pattern looks like GA morphology. Correlation study of the HURP p725 statuses and GA structure, site-directed mutagenesis and knockdown-rescue experiments were employed to identify the modified HURP as a key regulator assembling GA as a crescent ribbon.
Results
The cells containing no or extended distribution of HURP p725 have dispersed GA membranes or longer GA. Knockdown of HURP fragmentized GA and HURP wild type could, while its phosphorylation deficiency mutant 725A could not, restore crescent Golgi ribbon in HURP depleted cells, collectively indicating a crescent GA-constructing activity of HURP p725. HURP p725 is transported, by GA membrane-associated ARF1, Dynein and its cargo adaptor Golgin-160, to cell center where HURP p725 forms crescent fibers, binds and stabilizes Golgi assembly factors (GAFs) including TRIP11, GRASP65 and GM130, thereby dictating the formation of crescent Golgi ribbon at nuclear periphery.
Conclusions
The Ajuba/PRMT5/Aurora-A complex integrates the signals of protein methylation and phosphorylation to HURP, and the HURP p725 organizes GA by stabilizing and recruiting GAFs to its crescent-like structure, therefore sha** GA as a crescent ribbon. Therefore, the HURP p725 fiber serves a template to construct GA according to its shape.
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Background
The Golgi apparatus (GA) in mammalian cells forms a continuous ribbon of laterally interconnected stacks of flat cisternae. The construction of GA architecture relies on posttranslational modifications of soluble Golgi assembly factors (GAFs) and is subjected to dynamic changes along a cell cycle [2]. At G2 phase, phosphorylation-induced inactivation of GRASP65 [7] or GRASP55 [15], two GAFs engaged in stacking and linking cis- and trans-Golgi respectively, leads to the unlinking of the inter-connected stacks of the organelle. At early mitosis, further phosphorylation of the two GRASPs causes GA unstacking [41, The data used to support the findings of this study are included within the article. Golgi apparatus Golgi assembly factors Wild type Crescent ribbon shape Endoplasmic reticulum BFA Altan-Bonnet N, Phair RD, Polishchuk RS, Weigert R, Lippincott-Schwartz J. A role for Arf1 in mitotic Golgi disassembly, chromosome segregation, and cytokinesis. 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Ministry of Science and Technology (Taiwan), Taichung Veterans General Hospital (Taiwan), China Medical University (Taiwan), and China Medical Hospital (Taiwan). This work was supported by the grants from the Ministry of Science and Technology (MOST 108–2320-B-260–001, MOST 109–2314-B-039–047-MY3, 111–2320-B-260–001), the Taichung Veterans General Hospital-National Chi Nan University Joint Research Program (TCVGH-NCNU 1107903, TCVGH-NCNU 1097903), the China Medical University and Hospital grant (DMR-110–140) awarded to Dr. Shao-Chih Chiu, and the Taichung Veterans General Hospital (TCVGH-1093207D). Yu-Ting Amber Liao, **n-Ting Yang, Tong-You Wade Wei, and Jo-Mei Maureen Chen performed most of the experiments, Chun-Chih Jared Liu, Yu-Ting Jenny Huang, Yi-Chun Kuo, Chang-**n Wan, Chiao-Yun Cheng, Chen-Yu Chu and Yun-Ru Jaoying Huang assisted with some other experiments. Shao-Chih Chiu and Chang-Tze Ricky Yu supervised the study, and Chang-Tze Ricky Yu conceived and wrote the paper. The author(s) read and approved the final manuscript. Not applicable. (We did not perform experiment on animal or collect patient information). All authors have authorized that he/she will notify of his/her actions and the publication of this paper. The authors declare no competing interests. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Deduction of PRMT5-dependent methylation determinant sequence. Supplementary Figure 2. The HURP m122 antibodies and nm122 antibodies almost did not cross react. Supplementary Figure 3. Antibody specificity of HURP p725 and np725. 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The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. Chiu, SC., Yang, XT., Wei, TY.W. et al. The crescent-like Golgi ribbon is shaped by the Ajuba/PRMT5/Aurora-A complex-modified HURP.
Cell Commun Signal 21, 156 (2023). https://doi.org/10.1186/s12964-023-01167-4 Received: Accepted: Published: DOI: https://doi.org/10.1186/s12964-023-01167-4Availability of data and materials
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Additional file 1: Supplementary Figure 1.
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