TY - JOUR AU - Visnovitz, Tamás AU - Lenzinger, Dorina AU - Koncz, Anna AU - Vizi, Péter Márk AU - Bárkai, Tünde AU - Visnovitzné Dr Vukman, Krisztina AU - Galinsoga, Alicia AU - Németh, Krisztina AU - Fletcher, Kelsey Aine AU - Komlósi, Zsolt AU - Cserép, Csaba AU - Dénes, Ádám AU - Lőrincz, Péter AU - Valcz, Gábor AU - Buzás, Edit Irén TI - A “torn bag mechanism” of small extracellular vesicle release via limiting membrane rupture of en bloc released amphisomes (amphiectosomes) JF - ELIFE J2 - ELIFE VL - 13 PY - 2025 PG - 19 SN - 2050-084X DO - 10.7554/eLife.95828.3 UR - https://m2.mtmt.hu/api/publication/34720930 ID - 34720930 AB - Recent studies showed an unexpected complexity of extracellular vesicle (EV) biogenesis pathways. We previously found evidence that human colorectal cancer cells in vivo release large multivesicular body-like structures en bloc. Here, we tested whether this large EV type is unique to colorectal cancer cells. We found that all cell types we studied (including different cell lines and cells in their original tissue environment) released multivesicular large EVs (MV-lEVs). We also demonstrated that upon spontaneous rupture of the limiting membrane of the MV-lEVs, their intraluminal vesicles (ILVs) escaped to the extracellular environment by a ‘torn bag mechanism’. We proved that the MV-lEVs were released by ectocytosis of amphisomes (hence, we termed them amphiectosomes). Both ILVs of amphiectosomes and small EVs separated from conditioned media were either exclusively CD63 or LC3B positive. According to our model, upon fusion of multivesicular bodies with autophagosomes, fragments of the autophagosomal inner membrane curl up to form LC3B positive ILVs of amphisomes, while CD63 positive small EVs are of multivesicular body origin. Our data suggest a novel common release mechanism for small EVs, distinct from the exocytosis of multivesicular bodies or amphisomes, as well as the small ectosome release pathway. LA - English DB - MTMT ER - TY - JOUR AU - Boda, Attila AU - Balázs, Villő AU - Nagy, Anikó Zsuzsanna AU - Hargitai, Dávid AU - Lippai, Mónika AU - Simon-Vecsei, Zsófia Judit AU - Molnár, Márton AU - Fürstenhoffer, Fanni AU - Juhász, Gábor AU - Lőrincz, Péter TI - The Rab7-Epg5 and Rab39-ema modules cooperately position autophagosomes for efficient lysosomal fusions JF - ELIFE J2 - ELIFE VL - 13 PY - 2025 SN - 2050-084X DO - 10.7554/eLife.102663.1 UR - https://m2.mtmt.hu/api/publication/35447765 ID - 35447765 LA - English DB - MTMT ER - TY - GEN AU - Szenci, Győző AU - Boda, Attila AU - Nagy, Anikó Zsuzsanna AU - Szőke, Zsombor AU - Falcsik, Gergő AU - Kovács, Tibor AU - Lőrincz, Péter AU - Juhász, Gábor AU - Takáts, Szabolcs TI - Huntington’s disease-associated ankyrin repeat palmitoyl transferases are rate-limiting factors in lysosome formation and fusion PY - 2025 DO - 10.1101/2025.02.06.636816 UR - https://m2.mtmt.hu/api/publication/35755706 ID - 35755706 AB - Protein palmitoylation in the Golgi apparatus is critical for the appropriate sorting of various proteins belonging to secretory and lysosomal systems, and defective palmitoylation can lead to the onset of severe pathologies. HIP14 and HIP14L ankyrin repeat-containing palmitoyl transferases were linked to the pathogenesis of Huntington’s disease, however, how perturbation of these Golgi resident enzymes contributes to neurological disorders is yet to be understood. In this study, we investigated the function of Hip14 and Patsas - the Drosophila orthologs of HIP14 and HIP14L respectively – to uncover their role in secretory and lysosomal membrane trafficking. Using larval salivary gland, a well-established model of the regulated secretory pathway, we found that these PAT enzymes equally contribute to the proper maturation and crinophagic degradation of glue secretory granules by mediating their fusion with the endo-lysosomal compartment. We also revealed that Patsas and Hip14 are both required for lysosomal acidification and biosynthetic transport of various lysosomal hydrolases, and we demonstrated that the rate of secretory granule-lysosome fusion and subsequent acidification positively correlates with the level of Hip14. Furthermore, Hip14 is also essential for proper lysosome formation and neuronal function in adult brains. Finally, we found that the over-activation of lysosomal biosynthetic transport and lysosomal fusions by the expression of the constitutively active form of Rab2 could compensate for the lysosomal dysfunction caused by the loss of Patsas or Hip14 both in larval salivary glands and neurons. Therefore, we demonstrated that ankyrin repeat palmitoyl transferases may act as rate-limiting factors in lysosomal fusions and provide genetic evidence that defective protein palmitoylation and the subsequent lysosomal dysfunction can contribute to the onset of Huntington’s disease-like symptoms. LA - English DB - MTMT ER - TY - JOUR AU - Hargitai, Dávid AU - Nagy, Anikó Zsuzsanna AU - Bodor, Iván AU - Szenci, Győző AU - Laczkó-Dobos, Hajnalka AU - Bhattacharjee, Arindam AU - Neuhauser, Natali AU - Takáts, Szabolcs AU - Juhász, Gábor AU - Lőrincz, Péter TI - HOPS-dependent vesicle tethering lock inhibits endolysosomal fusions and autophagosome secretion upon the loss of Syntaxin17 JF - SCIENCE ADVANCES J2 - SCI ADV VL - 11 PY - 2025 IS - 23 SN - 2375-2548 DO - 10.1126/sciadv.adu9605 UR - https://m2.mtmt.hu/api/publication/36182547 ID - 36182547 N1 - Funding Agency and Grant Number: Hungarian Academy of Sciences [LP2022-13, LP2023-6, BO/00400/23]; National Research, Development, and innovation Office of Hungary [FK 138851, FK 142508, KKP129797]; Eotvos Lorand University Excellence Fund [EKA 2022/045-P101, EKA 2022/045-P302-1]; Ministry for Culture and innovation (NKP-23-3 New National Excellence Program and the DKOEP-23 Doctoral Excellence Program of the Ministry for Culture and innovation from the source of the National Research, Development and innovation Fund of Hunga [NKP-23-3-i-ELTE-724, DKOEP-2023-ELTE-13, NKP-23-5-ELTE-1257] Funding text: This work was funded by thefollowing organizations: Hungarian Academy of Sciences (MagyarTudomanyos Akademia): LP2022-13 to P.L., LP2023-6 to G.J. and BO/00400/23 to S.T.; National Research, Development, and innovation Office of Hungary (Nemzeti Kutatasi, Fejlesztesi es innovacios Hivatal): FK 138851 to P.L., FK 142508 to S.T. and Elvonal KKP129797 to G.J.; Eotvos Lorand University Excellence Fund: EKA 2022/045-P101 to P.L. and EKA 2022/045-P302-1 to S.T.; Ministry for Culture and innovation (& Uacute;NKP-23-3 New National Excellence Program and the DKOEP-23 Doctoral Excellence Program of the Ministry for Culture and innovation from the source of the National Research, Development and innovation Fund of Hungary): & Uacute;NKP-23-3-i-ELTE-724 and DKOEP-2023-ELTE-13 to D.H. and & Uacute;NKP-23-5-ELTE-1257 to S.T. AB - The autophagosomal SNARE (soluble N -ethylmaleimide–sensitive factor attachment protein) Syntaxin17 (Syx17) plays a pivotal role in autophagosome-lysosome fusion, yet the broader impact of its loss remains elusive. Our investigation of Syx17 function in Drosophila nephrocytes and salivary gland cells revealed unexpected effects. We find that Syx17 loss induces the formation of autophagosome-lysosome clusters in a HOPS (homotypic fusion and vacuole protein sorting)–dependent manner, entrapping this tether, autophagosomes, and lysosomes. While locked in clusters, these organelles cannot participate in other vesicle fusions, impeding endosomal progression and autophagosome secretion. Therefore, the absence of Syx17 not only inhibits autophagosome-lysosome fusion but also prevents HOPS release from autophagosome-lysosome tethering sites causing a “tethering lock.” Preventing autophagosome formation or removing the HOPS adaptor Plekhm1 (pleckstrin homology domain–containing family M member 1) leads to release of HOPS and lysosomes from these clusters, thus rescuing secondary effects of Syx17 loss. Our findings show that a tethering lock can disrupt multiple vesicle trafficking routes. LA - English DB - MTMT ER - TY - JOUR AU - Sőth, Ármin AU - Molnár, Márton AU - Lőrincz, Péter AU - Simon-Vecsei, Zsófia Judit AU - Juhász, Gábor TI - CORVET-specific subunit levels determine the balance between HOPS/CORVET endosomal tethering complexes JF - SCIENTIFIC REPORTS J2 - SCI REP VL - 14 PY - 2024 IS - 1 PG - 9 SN - 2045-2322 DO - 10.1038/s41598-024-59775-0 UR - https://m2.mtmt.hu/api/publication/34837906 ID - 34837906 N1 - Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University (ELTE), Pázmány Péter sétány 1/C, Budapest, 1117, Hungary Momentum Vesicle Trafficking Research Group, Hungarian Academy of Sciences-Eötvös Loránd University, Budapest, Hungary Momentum Lysosomal Degradation Research Group, Institute of Genetics, HUN-REN Biological Research Centre Szeged, Szeged, Hungary Export Date: 6 June 2024 Correspondence Address: Simon-Vecsei, Z.; Department of Anatomy, Pázmány Péter sétány 1/C, Hungary; email: simon.vecsei.zsofia@ttk.elte.hu Correspondence Address: Juhász, G.; Department of Anatomy, Pázmány Péter sétány 1/C, Hungary; email: gabor.juhasz@ttk.elte.hu AB - The closely related endolysosomal tethering complexes HOPS and CORVET play pivotal roles in the homo- and heterotypic fusion of early and late endosomes, respectively, and HOPS also mediates the fusion of lysosomes with incoming vesicles including late endosomes and autophagosomes. These heterohexameric complexes share their four core subunits that assemble with additional two, complex-specific subunits. These features and the similar structure of the complexes could allow the formation of hybrid complexes, and the complex specific subunits may compete for binding to the core. Indeed, our biochemical analyses revealed the overlap of binding sites for HOPS-specific VPS41 and CORVET-specific VPS8 on the shared core subunit VPS18. We found that the overexpression of CORVET-specific VPS8 or Tgfbrap1 decreased the amount of core proteins VPS11 and VPS18 that are assembled with HOPS-specific subunits VPS41 or VPS39, indicating reduced amount of assembled HOPS. In line with this, we observed the elevation of both lipidated, autophagosome-associated LC3 protein and the autophagic cargo p62 in these cells, suggesting impaired autophagosome-lysosome fusion. In contrast, overexpression of HOPS-specific VPS39 or VPS41 did not affect the level of assembled CORVET or autophagy. VPS8 or Tgfbrap1 overexpression also induced Cathepsin D accumulation, suggesting that HOPS-dependent biosynthetic delivery of lysosomal hydrolases is perturbed, too. These indicate that CORVET-specific subunit levels fine-tune HOPS assembly and activity in vivo. LA - English DB - MTMT ER - TY - JOUR AU - Boda, Attila AU - Varga, Luca Petra AU - Nagy, Anikó Zsuzsanna AU - Szenci, Győző AU - Csizmadia, Tamás AU - Lőrincz, Péter AU - Juhász, Gábor TI - Rab26 controls secretory granule maturation and breakdown in Drosophila JF - CELLULAR AND MOLECULAR LIFE SCIENCES J2 - CELL MOL LIFE SCI VL - 80 PY - 2023 IS - 1 PG - 15 SN - 1420-682X DO - 10.1007/s00018-022-04674-8 UR - https://m2.mtmt.hu/api/publication/33542161 ID - 33542161 N1 - Export Date: 17 January 2023 CODEN: CMLSF AB - At the onset of Drosophila metamorphosis, plenty of secretory glue granules are released from salivary gland cells and the glue is deposited on the ventral side of the forming (pre)pupa to attach it to a dry surface. Prior to this, a poorly understood maturation process takes place during which secretory granules gradually grow via homotypic fusions, and their contents are reorganized. Here we show that the small GTPase Rab26 localizes to immature (smaller, non-acidic) glue granules and its presence prevents vesicle acidification. Rab26 mutation accelerates the maturation, acidification and release of these secretory vesicles as well as the lysosomal breakdown (crinophagy) of residual, non-released glue granules. Strikingly, loss of Mon1, an activator of the late endosomal and lysosomal fusion factor Rab7, results in Rab26 remaining associated even with the large glue granules and a concomitant defect in glue release, similar to the effects of Rab26 overexpression. Our data thus identify Rab26 as a key regulator of secretory vesicle maturation that promotes early steps (vesicle growth) and inhibits later steps (lysosomal transport, acidification, content reorganization, release, and breakdown), which is counteracted by Mon1. LA - English DB - MTMT ER - TY - JOUR AU - Rzepiel, Andrea AU - Horváth, Anna AU - Kutszegi, Nóra AU - Gézsi, András AU - C. Sági, Judit AU - Almási, Laura AU - Egyed, Bálint AU - Lőrincz, Péter AU - Visnovitz, Tamás AU - Kovács, Gábor AU - Szalai, Csaba AU - Semsei F, Ágnes AU - Erdélyi, Dániel TI - MiR-128-3p as blood based liquid biopsy biomarker in childhood acute lymphoblastic leukemia JF - MOLECULAR AND CELLULAR PROBES J2 - MOL CELL PROBE VL - 67 PY - 2023 PG - 7 SN - 0890-8508 DO - 10.1016/j.mcp.2023.101893 UR - https://m2.mtmt.hu/api/publication/33577132 ID - 33577132 N1 - Dept. of Paediatrics, Semmelweis University, Budapest, Hungary Dept. of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, Hungary Dept. of Measurement and Information Systems, Budapest University of Technology and Economics, Budapest, Hungary Institute of Genomic Medicine and Rare Disorders, Semmelweis University, Budapest, Hungary HCEMM-SE Molecular Oncohematology Research Group, 1st Dept. of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary Dept. of Anatomy, Cell and Developmental Biology, ELTE Eötvös Loránd University, Budapest, Hungary Dept. of Plant Physiology and Molecular Plant Biology, ELTE Eötvös Loránd University, Budapest, Hungary Heim Pál Children Hospital, Budapest, Hungary Cited By :1 Export Date: 24 April 2023 CODEN: MCPRE Correspondence Address: Semsei, Á.F.; Dept. of Genetics, 1085 Üllői út 26, Hungary; email: semsei.agnes@med.semmelweis-univ.hu LA - English DB - MTMT ER - TY - JOUR AU - Koncz, Anna AU - Turiák, Lilla AU - Németh, Krisztina AU - Lenzinger, Dorina AU - Bárkai, Tünde AU - Lőrincz, Péter AU - Zelenyánszki, Helga AU - Visnovitzné Dr Vukman, Krisztina AU - Buzás, Edit Irén AU - Visnovitz, Tamás TI - Endoplasmin Is a Hypoxia-Inducible Endoplasmic Reticulum-Derived Cargo of Extracellular Vesicles Released by Cardiac Cell Lines JF - MEMBRANES (BASEL) J2 - MEMBRANES-BASEL VL - 13 PY - 2023 IS - 4 PG - 16 SN - 2077-0375 DO - 10.3390/membranes13040431 UR - https://m2.mtmt.hu/api/publication/33750753 ID - 33750753 N1 - Department of Genetics, Cell and Immunobiology, Semmelweis University, Üllői út 26, Budapest, 1085, Hungary Research Centre for Natural Sciences, Institute of Organic Chemistry, Magyar Tudósok Körútja 2, Budapest, 1117, Hungary ELKH-SE Translational Extracellular Vesicle Research Group, Nagyvárad tér 4, Budapest, 1085, Hungary Department of Anatomy, Cell and Developmental Biology, ELTE Eötvös Loránd University, Pázmány Péter sétány 1/c, Budapest, 1117, Hungary Department of Plant Physiology and Molecular Plant Biology, ELTE Eötvös Loránd University, Pázmány Péter sétány 1/c, Budapest, 1117, Hungary HCEMM-SU Extracellular Vesicle Research Group, Nagyvárad tér 4, Budapest, 1085, Hungary Export Date: 30 May 2023 Correspondence Address: Visnovitz, T.; Department of Genetics, Üllői út 26, Hungary; email: visnovitz.tamas@med.semmelweis-univ.hu AB - Cardiomyopathies are leading causes of human mortality. Recent data indicate that the cardiomyocyte-derived extracellular vesicles (EVs) released upon cardiac injury are present in circulation. This paper aimed to analyze EVs released under normal and hypoxic conditions by H9c2 (rat), AC16 (human) and HL1 (mouse) cardiac cell lines. Small (sEVs), medium (mEVs) and large EVs (lEVs) were separated from a conditioned medium by a combination of gravity filtration, differential centrifugation and tangential flow filtration. The EVs were characterized by microBCA, SPV lipid assay, nanoparticle tracking analysis, transmission and immunogold electron microscopy, flow cytometry and Western blotting. Proteomic profiles of the EVs were determined. Surprisingly, an endoplasmic reticulum chaperone, endoplasmin (ENPL, grp94 or gp96), was identified in the EV samples, and its association with EVs was validated. The secretion and uptake of ENPL was followed by confocal microscopy using GFP-ENPL fusion protein expressing HL1 cells. We identified ENPL as an internal cargo of cardiomyocyte-derived mEVs and sEVs. Based on our proteomic analysis, its presence in EVs was linked to hypoxia in HL1 and H9c2 cells, and we hypothesize that EV-associated ENPL may have a cardioprotective role by reducing cardiomyocyte ER stress. LA - English DB - MTMT ER - TY - JOUR AU - Maruzs, Tamás AU - Feil-Börcsök, Dalma AU - Lakatos, Enikő AU - Juhász, Gábor AU - Blastyák, András AU - Hargitai, Dávid AU - Jean, Steve AU - Lőrincz, Péter AU - Juhász, Gábor TI - Interaction of the sorting nexin 25 homologue Snazarus with Rab11 balances endocytic and secretory transport and maintains the ultrafiltration diaphragm in nephrocytes JF - MOLECULAR BIOLOGY OF THE CELL J2 - MOL BIOL CELL VL - 34 PY - 2023 IS - 9 PG - 14 SN - 1059-1524 DO - 10.1091/mbc.E22-09-0421 UR - https://m2.mtmt.hu/api/publication/34021340 ID - 34021340 N1 - Funding Agency and Grant Number: National Research Development and Innovation Office (NKFIH) of Hungary [PD135611, UNKP-22-2-III-ELTE-702, FK138851, Elvonal KKP129797]; National Laboratory of Biotechnology [2022-2.1.1-NL-2022-00008]; Eotvos Lorand University Excellence Fund [EKA 2022/045-P101-2]; National Academy of Scientist Education under the sponsorship of the Hungarian Ministry of Innovation and Technology [FEIF/646-4/2021-ITM_SZERZ] Funding text: We thank Szilvia Bozso and Monika Truszka for technical assistance. We thank Mike W. Henne for suggestions and for providing reagents and Michael Krahn and Thomas L. Schwarz for providing reagents. This work was supported by the National Research Development and Innovation Office (NKFIH) of Hungary with PD135611 grant to T.M., UNKP-22-2-III-ELTE-702 grant to D.H., FK138851 grant to P.L., and Elvonal KKP129797 and the National Laboratory of Biotechnology 2022-2.1.1-NL-2022-00008 grants to G.J. The work was also supported by the Eotvos Lorand University Excellence Fund (EKA 2022/045-P101-2) to P.L.. This research work was conducted with additional support from the National Academy of Scientist Education under the sponsorship of the Hungarian Ministry of Innovation and Technology (FEIF/646-4/2021-ITM_SZERZ). AB - Proper balance of exocytosis and endocytosis is important for the maintenance of plasma membrane lipid and protein homeostasis. This is especially critical in human podocytes and the podocyte-like Drosophila nephrocytes that both use a delicate diaphragm system with evolutionarily conserved components for ultrafiltration. Here we show that the sorting nexin 25 homolog Snazarus (Snz) binds to Rab11 and localizes to Rab11-positive recycling endosomes in Drosophila nephrocytes, unlike in fat cells where it is present in plasma membrane/lipid droplet/ER contact sites. Loss of Snz leads to redistribution of Rab11 vesicles from the cell periphery and increases endocytic activity in nephrocytes. These changes are accompanied by defects in diaphragm protein distribution that resemble those seen in Rab11 gain-of-function cells. Of note, co-overexpression of Snz rescues diaphragm defects in Rab11 overexpressing cells, whereas snz knockdown in Rab11 overexpressing nephrocytes or simultaneous knockdown of snz and tbc1d8b encoding a Rab11 GAP lead to massive expansion of the lacunar system that contains mislocalized diaphragm components: Sns and Pyd/ZO-1. We find that loss of Snz enhances while its overexpression impairs secretion, which, together with genetic epistasis analyses, suggest that Snz counteracts Rab11 to maintain the diaphragm via setting the proper balance of exocytosis and endocytosis. LA - English DB - MTMT ER - TY - JOUR AU - Vedelek, Viktor AU - Vedelek, Balázs AU - Lőrincz, Péter AU - Juhász, Gábor AU - Sinka, Rita TI - A comparative analysis of fruit fly and human glutamate dehydrogenases in Drosophila melanogaster sperm development JF - FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY J2 - FRONT CELL DEV BIOL VL - 11 PY - 2023 PG - 16 SN - 2296-634X DO - 10.3389/fcell.2023.1281487 UR - https://m2.mtmt.hu/api/publication/34239207 ID - 34239207 N1 - Funding Agency and Grant Number: The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by NKFIH (PD137914 to VV, K132155 to RS, and FK138851 to PL), National Research, Development, and Innovation Offi [PD137914, K132155, FK138851]; NKFIH [2022-2.1.1-NL-2022-00008, EKA 2022/045-P101-2, LP 2022-13/2022]; National Research, Development, and Innovation Office of Hungary (Biotechnology National Laboratory) Funding text: The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by NKFIH (PD137914 to VV, K132155 to RS, and FK138851 to PL), National Research, Development, and Innovation Office of Hungary (Biotechnology National Laboratory 2022-2.1.1-NL-2022-00008 to GJ), and Eotvos Lorand University Excellence Fund (EKA 2022/045-P101-2), MTA-t (LP 2022-13/2022) to PL.r The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by NKFIH (PD137914 to VV, K132155 to RS, and FK138851 to PL), National Research, Development, and Innovation Office of Hungary (Biotechnology National Laboratory 2022-2.1.1-NL-2022-00008 to GJ), and Eotvos Lorand University Excellence Fund (EKA 2022/045-P101-2), MTA-t (LP 2022-13/2022) to PL. AB - Glutamate dehydrogenases are enzymes that take part in both amino acid and energy metabolism. Their role is clear in many biological processes, from neuronal function to cancer development. The putative testis-specific Drosophila glutamate dehydrogenase, Bb8, is required for male fertility and the development of mitochondrial derivatives in spermatids. Testis-specific genes are less conserved and could gain new functions, thus raising a question whether Bb8 has retained its original enzymatic activity. We show that while Bb8 displays glutamate dehydrogenase activity, there are significant functional differences between the housekeeping Gdh and the testis-specific Bb8. Both human GLUD1 and GLUD2 can rescue the bb8 ms mutant phenotype, with superior performance by GLUD2. We also tested the role of three conserved amino acids observed in both Bb8 and GLUD2 in Gdh mutants, which showed their importance in the glutamate dehydrogenase function. The findings of our study indicate that Drosophila Bb8 and human GLUD2 could be novel examples of convergent molecular evolution. Furthermore, we investigated the importance of glutamate levels in mitochondrial homeostasis during spermatogenesis by ectopic expression of the mitochondrial glutamate transporter Aralar1, which caused mitochondrial abnormalities in fly spermatids. The data presented in our study offer evidence supporting the significant involvement of glutamate metabolism in sperm development. LA - English DB - MTMT ER -