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 PY - 2024 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 - 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 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 - Visnovitz, Tamás AU - Lenzinger, Dorina AU - Koncz, Anna AU - Vizi, Péter M AU - Bárkai, Tünde AU - Visnovitzné Dr Vukman, Krisztina AU - Galinsoga, Alicia AU - Németh, Krisztina AU - Fletcher, Kelsey AU - Komlósi, Zsolt 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 - 2024 PG - 35 SN - 2050-084X DO - 10.7554/eLife.95828.1 UR - https://m2.mtmt.hu/api/publication/34720930 ID - 34720930 LA - English DB - MTMT ER - TY - CONF AU - Vedelek, Viktor AU - Balázs, Vedelek AU - Gabor, Juhasz AU - Lőrincz, Péter AU - Sinka, Rita TI - Convergent evolution in glutamate dehydrogenase activity in Drosophila and human T2 - 27th European Drosophila Research Conference PY - 2023 PG - 1 UR - https://m2.mtmt.hu/api/publication/34684460 ID - 34684460 LA - English DB - MTMT ER - TY - GEN AU - Kovács, Kinga Dóra AU - Visnovitz, Tamás AU - Kanyó, Nicolett AU - Gerecsei, Tamás AU - Péter, Beatrix AU - Lagzi, István AU - Kurunczi, Sándor AU - Koncz, Anna AU - Németh, Krisztina AU - Lenzinger, Dorina AU - V. Vukman, Krisztina AU - Molnár, Kinga AU - Truszka, Mónika AU - Nakanishi, Hideyuki AU - Lőrincz, Péter AU - Székács, Inna AU - Buzás, Edit I. AU - Horváth, Róbert TI - Nanoinjection of fluorescent and gold nanoparticles to single live cells by robotic fluidic force microscopy PY - 2023 UR - https://m2.mtmt.hu/api/publication/34486423 ID - 34486423 LA - English DB - MTMT ER - TY - CHAP AU - Kovács, Kinga Dóra AU - Visnovitz, Tamás AU - Gerecsei, Tamás AU - Péter, Beatrix AU - Kurunczi, Sándor AU - Koncz, Anna AU - Németh, Krisztina AU - Lenzinger, Dorina AU - V. Vukman, Krisztina AU - Lőrincz, Péter AU - Székács, Inna AU - Buzás, Edit I. AU - Horváth, Róbert ED - Derényi, Imre ED - László, Grama ED - Solymosi, Katalin TI - Nanoinjection of fluorescent nanoparticles to single live cells by robotic fluidic force microscopy T2 - Magyar Biofizikai Társaság XXIX. Kongresszusa PB - Magyar Biofizikai Társaság CY - Budapest SN - 9786150186887 PY - 2023 SP - 35 EP - 35 PG - 1 UR - https://m2.mtmt.hu/api/publication/34486404 ID - 34486404 LA - English DB - MTMT ER - TY - JOUR AU - Kovács, Kinga Dóra AU - Visnovitz, Tamás AU - Gerecsei, Tamás AU - Péter, Beatrix AU - Kurunczi, Sándor AU - Koncz, Anna AU - Németh, Krisztina AU - Lenzinger, Dorina AU - Visnovitzné Dr Vukman, Krisztina AU - Balogh, Anna AU - Rajmon, Imola AU - Lőrincz, Péter AU - Székács, Inna AU - Buzás, Edit Irén AU - Horváth, Róbert TI - Nanoinjection of extracellular vesicles to single live cells by robotic fluidic force microscopy JF - JOURNAL OF EXTRACELLULAR VESICLES J2 - J EXTRACELLULAR VESICL VL - 12 PY - 2023 IS - 12 PG - 16 SN - 2001-3078 DO - 10.1002/jev2.12388 UR - https://m2.mtmt.hu/api/publication/34425078 ID - 34425078 AB - In the past decade, extracellular vesicles (EVs) have attracted substantial interest in biomedicine. With progress in the field, we have an increasing understanding of cellular responses to EVs. In this Technical Report, we describe the direct nanoinjection of EVs into the cytoplasm of single cells of different cell lines. By using robotic fluidic force microscopy (robotic FluidFM), nanoinjection of GFP positive EVs and EV‐like particles into single live HeLa, H9c2, MDA‐MB‐231 and LCLC‐103H cells proved to be feasible. This injection platform offered the advantage of high cell selectivity and efficiency. The nanoinjected EVs were initially localized in concentrated spot‐like regions within the cytoplasm. Later, they were transported towards the periphery of the cells. Based on our proof‐of‐principle data, robotic FluidFM is suitable for targeting single living cells by EVs and may lead to information about intracellular EV cargo delivery at a single‐cell level. 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 - 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 - 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 -