TY - CONF AU - Páhi, Zoltán Gábor AU - Kovács, Levente AU - Szűcs, Diána AU - Borsos, Barbara Nikolett AU - Deák, Péter AU - Pankotai, Tibor TI - Usp5, Usp34, and Otu1 deubiquitylases mediate DNA repair in Drosophila melanogaster T2 - Ubiquitin & Friends Symposium 2023 PB - University of Vienna C1 - Bécs PY - 2023 SP - 67 EP - 67 PG - 1 UR - https://m2.mtmt.hu/api/publication/34506866 ID - 34506866 LA - English DB - MTMT ER - TY - JOUR AU - Farkas, Anita AU - Zsindely, Nóra AU - Nagy, Gábor AU - Kovács, Levente AU - Deák, Péter AU - Bodai, László TI - The ubiquitin thioesterase YOD1 ameliorates mutant Huntingtin induced pathology in Drosophila JF - SCIENTIFIC REPORTS J2 - SCI REP VL - 13 PY - 2023 IS - 1 PG - 13 SN - 2045-2322 DO - 10.1038/s41598-023-49241-8 UR - https://m2.mtmt.hu/api/publication/34431874 ID - 34431874 AB - Huntington’s disease (HD) is a neurodegenerative disorder caused by a dominant gain-of-function mutation in the huntingtin gene, resulting in an elongated polyglutamine repeat in the mutant Huntingtin (mHtt) that mediates aberrant protein interactions. Previous studies implicated the ubiquitin–proteasome system in HD, suggesting that restoring cellular proteostasis might be a key element in suppressing pathology. We applied genetic interaction tests in a Drosophila model to ask whether modulating the levels of deubiquitinase enzymes affect HD pathology. By testing 32 deubiquitinase genes we found that overexpression of Yod1 ameliorated all analyzed phenotypes, including neurodegeneration, motor activity, viability, and longevity. Yod1 did not have a similar effect in amyloid beta overexpressing flies, suggesting that the observed effects might be specific to mHtt. Yod1 overexpression did not alter the number of mHtt aggregates but moderately increased the ratio of larger aggregates. Transcriptome analysis showed that Yod1 suppressed the transcriptional effects of mHtt and restored the expression of genes involved in neuronal plasticity, vesicular transport, antimicrobial defense, and protein synthesis, modifications, and clearance. Furthermore, Yod1 overexpression in HD flies leads to the upregulation of genes involved in transcriptional regulation and synaptic transmission, which might be part of a response mechanism to mHtt-induced stress. LA - English DB - MTMT ER - TY - JOUR AU - Kovács, Levente AU - Fatalska, Agnieszka AU - Glover, David M. TI - Targeting Drosophila Sas6 to mitochondria reveals its high affinity for Gorab JF - BIOLOGY OPEN J2 - BIOL OPEN VL - 11 PY - 2022 IS - 11 PG - 8 SN - 2046-6390 DO - 10.1242/bio.059545 UR - https://m2.mtmt.hu/api/publication/33833753 ID - 33833753 AB - The ability to relocalize proteins to defined subcellular locations presents a powerful tool to examine protein-protein interactions that can overcome a tendency of non-targeted exogenous proteins to form inaccessible aggregates. Here, we show that a 24-amino-acid sequence from the Drosophila proapoptotic protein Hid's tail anchor (HTA) domain can target exogenous proteins to the mitochondria in Drosophila cells. We use this HTA tag to target the Drosophila centriole cartwheel protein Sas6 to the mitochondria, and show that both exogenous and endogenous Gorab can be co-recruited from the Golgi to the new mitochondrial site. This accords with our previous observation that monomeric Drosophila Gorab binds Sas6 to become centriole associated with a 50-fold greater affinity than dimeric Gorab binds Rab6 to become localized at the Golgi. Strikingly, Drosophila Sas6 can bind both Drosophila Gorab and its human GORAB ortholog, whereas human SAS6 is unable to bind either GORAB or Gorab. We discuss these findings in relation to the evolutionary conservation of Gorab and the divergence of Sas6, possibly reflecting known differences in persistence of the cartwheel in the centriole duplication cycle of fly and human cells. LA - English DB - MTMT ER - TY - JOUR AU - Páhi, Zoltán Gábor AU - Kovács, Levente AU - Szűcs, Diána AU - Borsos, Barbara Nikolett AU - Deák, Péter AU - Pankotai, Tibor TI - Usp5, Usp34, and Otu1 deubiquitylases mediate DNA repair in Drosophila melanogaster JF - SCIENTIFIC REPORTS J2 - SCI REP VL - 12 PY - 2022 IS - 1 PG - 11 SN - 2045-2322 DO - 10.1038/s41598-022-09703-x UR - https://m2.mtmt.hu/api/publication/32778093 ID - 32778093 N1 - Institute of Pathology, Albert Szent-Györgyi Medical School, University of Szeged, 1 Állomás Street, Szeged, 6725, Hungary Department of Genetics, Faculty of Sciences and Informatics, University of Szeged, 52 Közép fasor, Szeged, 6726, Hungary Divison of Biology and Biological Engineering, California Institute of Tehnology, 1200 East California Boulevard, Pasadena, 91125, United States Cited By :2 Export Date: 29 February 2024 Correspondence Address: Pankotai, T.; Institute of Pathology, 1 Állomás Street, Hungary; email: pankotai.tibor@szte.hu Chemicals/CAS: hydrolase, 9027-41-2; ubiquitin, 60267-61-0; Drosophila Proteins; Hydrolases; Ubiquitin; Ubiquitin-Specific Proteases; USP5 protein, Drosophila Funding details: Magyar Tudományos Akadémia, MTA, BO/27/20, ÚNKP-20-5-SZTE-265, ÚNKP-21-5-SZTE-563 Funding details: National Research, Development and Innovation Office, GINOP-2.2.1-15-2017-00052, GINOP-2.3.2-15-2016-00032, NKFI-FK 132080 Funding text 1: Open access funding provided by University of Szeged. This research was funded by National Research, Development and Innovation Office Grant GINOP-2.2.1-15-2017-00052, GINOP-2.3.2-15-2016-00032 and NKFI-FK 132080, the János Bolyai Research Scholarship of the Hungarian Academy of Sciences BO/27/20, ÚNKP-20-5-SZTE-265 and ÚNKP-21-5-SZTE-563. LA - English DB - MTMT ER - TY - JOUR AU - Bhattacharjee, Arindam AU - Harmatos-Ürmösi, Adél AU - Jipa, András AU - Kovács, Levente AU - Deák, Péter AU - Szabó, Áron AU - Juhász, Gábor TI - Loss of ubiquitinated protein autophagy is compensated by persistent cnc/NFE2L2/Nrf2 antioxidant responses JF - AUTOPHAGY J2 - AUTOPHAGY VL - 18 PY - 2022 IS - 10 SP - 2385 EP - 2396 PG - 12 SN - 1554-8627 DO - 10.1080/15548627.2022.2037852 UR - https://m2.mtmt.hu/api/publication/32691300 ID - 32691300 N1 - Funding Agency and Grant Number: National Research, Development and Innovation OfficeNational Research, Development & Innovation Office (NRDIO) - Hungary [KKP129797, GINOP-2.3.2-15-2016-00032, FK132183]; National Research, Development and Innovation Office[National Laboratory for Biotechnology program] [PD135587]; Hungarian Academy of SciencesHungarian Academy of Sciences [LP-2014/2, BO/00078/18, UNKP-20-5, UNKP-19-4] Funding text: This work was supported by the National Research, Development and Innovation Office[KKP129797; GINOP-2.3.2-15-2016-00032; FK132183; National Laboratory for Biotechnology program; PD135587] and the Hungarian Academy of Sciences [LP-2014/2; BO/00078/18; UNKP-20-5; UNKP-19-4]. LA - English DB - MTMT ER - TY - CHAP AU - Anna, Dósa AU - Csizmadia, Tamás AU - Maddali, Asha Kiran AU - Laczkó-Dobos, Hajnalka AU - Jipa, András AU - Kovács, Levente AU - Deák, Péter AU - Lőw, Péter AU - Juhász, Gábor ED - László, Buday ED - Miklós, Erdélyi ED - Beáta, Lontay ED - József, Mihály ED - Sinka, Rita ED - László, Virág ED - Gergely, Szakáts ED - Attila, Varga TI - The role of E3 ubiquitin ligases in the regulation of crinophagy T2 - Hungarian Molecular Life Sciences 2021 PB - Diamond Congress Kft. CY - Eger SN - 9786155270673 PY - 2021 SP - 1 UR - https://m2.mtmt.hu/api/publication/32576298 ID - 32576298 AB - Crinophagy is a special form of autophagy, in which damaged or no longer needed secretory granules directly fuse with lysosomes, independently from autophagosome formation. Our colleagues have previously identified the molecular components of the membrane fusion apparatus (SNARE proteins, small GTPases and the tethering complex), using Drosophila melanogaster as a model organism, in which crinophagy is part of the normal developmental program of the larval salivary gland. Despite these findings, our knowledge is still little about the complete mechanism, as the upstream regulation of the process remains mainly unclear. The well-known function of ubiquitin is the targeting of cytoplasmic proteins into the proteasome for degradation, maintaining normal protein turnover. However, it has been observed that ubiquitin plays important roles in many other cellular processes, such as signal transduction or proteasome-independent degradation. It has been previously shown that in yeast, an E3 ubiquitin ligase activity is needed for the fast degradation of the gluconeogenic enzymes through the Vacuolar Import and Degradation (Vid) pathway, which is mechanistically similar to crinophagy. According to these data, ubiquitin could potentially have a role in the regulation of crinophagy, too, so our group decided to investigate this possibility. In our work, using fluorescent and immunocytochemical labeling methods, we have found ubiquitin on the surface of (glue) secretory granules, when crinophagy is activated. Moreover, through a small- scale genetic screen that involved the genes of E3 ubiquitin ligases, we identified one E3 enzyme which is required for the initiation of developmentally programmed crinophagy. Importantly, the overexpression of this enzyme causes early induction of secretory granule-lysosome fusion. We observed these phenotypes using both fluorescent and electron microscopy techniques. According to these results, E3 ubiquitin ligase activity is likely needed for successful crinophagy, thus we predict that at least one ubiquitination event occurs in the process of crinophagy LA - English DB - MTMT ER - TY - CHAP AU - Csizmadia, Tamás AU - Dósa, Anna AU - Maddali, Asha Kiran AU - Laczkó-Dobos, Hajnalka AU - Jipa, András AU - Kovács, Levente AU - Deák, Péter AU - Lőw, Péter AU - Juhász, Gábor ED - László, Buday ED - Miklós, Erdélyi ED - Beáta, Lontay ED - József, Mihály ED - Sinka, Rita ED - László, Virág ED - Gergely, Szakáts ED - Attila, Varga TI - The role of ubiquitin in the secretory granule degradation in Drosophila T2 - Hungarian Molecular Life Sciences 2021 PB - Diamond Congress Kft. CY - Eger SN - 9786155270673 PY - 2021 SP - 0 UR - https://m2.mtmt.hu/api/publication/32576272 ID - 32576272 AB - In the late larval period of Drosophila salivary gland, part of the glue containing secretory granules evade exocytosis and are degraded via crinophagy. During this process the obsolete secretory vesicles directly fuse with late endosomes or lysosomes for the fast degradation and recycling of the secretory cargo. The mechanism of secretory granule assortment for degradation is unknown since the discovery of crinophagy. Here we show that, the obsolete secretory granules are designated for degradation by ubiquitin (Figure 1) before their fusion with late endosomes/lysosomes. Moreover, we have identified the molecular components (E3 and deubiquitinating enzymes), which are involved in the ubiquitination of secretory (glue) granule membrane proteins. Importantly, the activity of these components is required for the normal crinophagic degradation of unnecessary secretory vesicles. Our work establishes the molecular background, which directs obsolete secretory granules for developmentally programmed crinophagy in Drosophila and paves the way for analyzing and understanding the regulation of this process in Metazoan LA - English DB - MTMT ER - TY - JOUR AU - Fatalska, Agnieszka AU - Stepinac, Emma AU - Richter, Magdalena AU - Kovács, Levente AU - Pietras, Zbigniew AU - Puchinger, Martin AU - Dong, Gang AU - Dadlez, Michal AU - Glover, David M. TI - The dimeric Golgi protein Gorab binds to Sas6 as a monomer to mediate centriole duplication JF - ELIFE J2 - ELIFE VL - 10 PY - 2021 PG - 24 SN - 2050-084X DO - 10.7554/eLife.57241 UR - https://m2.mtmt.hu/api/publication/32367729 ID - 32367729 N1 - Department of Genetics, University of Cambridge, Cambridge, United Kingdom Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, United States Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland Department of Medical Biochemistry, Max Perutz Labs, Medical University of Vienna, Vienna, Austria Department of Structural and Computational Biology, Max Perutz Labs, University of Vienna, Vienna, Austria Export Date: 30 November 2021 Correspondence Address: Fatalska, A.; Department of Genetics, United Kingdom; email: af589@cam.ac.uk Correspondence Address: Fatalska, A.; Division of Biology and Biological Engineering, United States; email: af589@cam.ac.uk Correspondence Address: Fatalska, A.; Institute of Biochemistry and Biophysics, Poland; email: af589@cam.ac.uk Correspondence Address: Glover, D.M.; Department of Genetics, United Kingdom; email: dmglover@caltech.edu Correspondence Address: Glover, D.M.; Division of Biology and Biological Engineering, United States; email: dmglover@caltech.edu AB - The duplication and ninefold symmetry of the Drosophila centriole requires that the cartwheel molecule, Sas6, physically associates with Gorab, a trans-Golgi component. How Gorab achieves these disparate associations is unclear. Here, we use hydrogen-deuterium exchange mass spectrometry to define Gorab's interacting surfaces that mediate its subcellular localization. We identify a core stabilization sequence within Gorab's C-terminal coiled-coil domain that enables homodimerization, binding to Rab6, and thereby trans-Golgi localization. By contrast, part of the Gorab monomer's coiled-coil domain undergoes an antiparallel interaction with a segment of the parallel coiled-coil dimer of Sas6. This stable heterotrimeric complex can be visualized by electron microscopy. Mutation of a single leucine residue in Sas6's Gorab-binding domain generates a Sas6 variant with a sixteenfold reduced binding affinity for Gorab that cannot support centriole duplication. Thus, Gorab dimers at the Golgi exist in equilibrium with Sas6-associated monomers at the centriole to balance Gorab's dual role. LA - English DB - MTMT ER - TY - JOUR AU - Panda, P. AU - Kovács, Levente AU - Dzhindzhev, N. AU - Fatalska, A. AU - Persico, V. AU - Geymonat, M. AU - Riparbelli, M.G. AU - Callaini, G. AU - Glover, D.M. TI - Tissue specific requirement of Drosophila Rcd4 for centriole duplication and ciliogenesis JF - JOURNAL OF CELL BIOLOGY J2 - J CELL BIOL VL - 219 PY - 2020 IS - 8 PG - 23 SN - 0021-9525 DO - 10.1083/jcb.201912154 UR - https://m2.mtmt.hu/api/publication/31383527 ID - 31383527 N1 - Department of Genetics, University of Cambridge, Cambridge, United Kingdom Institute of Biochemistry and Biophysics, Polish Academy of Science, Warsaw, Poland Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, United States Department of Life Sciences, University of Siena, Siena, Italy Export Date: 20 July 2020 CODEN: JCLBA Correspondence Address: Glover, D.M.; Department of Genetics, University of CambridgeUnited Kingdom; email: dmglover@caltech.edu Department of Genetics, University of Cambridge, Cambridge, United Kingdom Institute of Biochemistry and Biophysics, Polish Academy of Science, Warsaw, Poland Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, United States Department of Life Sciences, University of Siena, Siena, Italy Export Date: 13 January 2021 CODEN: JCLBA Correspondence Address: Glover, D.M.; Department of Genetics, University of CambridgeUnited Kingdom; email: dmglover@caltech.edu AB - Rcd4 is a poorly characterized Drosophila centriole component whose mammalian counterpart, PPP1R35, is suggested to function in centriole elongation and conversion to centrosomes. Here, we show that rcd4 mutants exhibit fewer centrioles, aberrant mitoses, and reduced basal bodies in sensory organs. Rcd4 interacts with the C-terminal part of Ana3, which loads onto the procentriole during interphase, ahead of Rcd4 and before mitosis. Accordingly, depletion of Ana3 prevents Rcd4 recruitment but not vice versa. We find that neither Ana3 nor Rcd4 participates directly in the mitotic conversion of centrioles to centrosomes, but both are required to load Ana1, which is essential for such conversion. Whereas ana3 mutants are male sterile, reflecting a requirement for Ana3 for centriole development in the male germ line, rcd4 mutants are fertile and have male germ line centrioles of normal length. Thus, Rcd4 is essential in somatic cells but is not absolutely required in spermatogenesis, indicating tissue-specific roles in centriole and basal body formation. © 2020 Panda et al. LA - English DB - MTMT ER - TY - JOUR AU - Kovács, Levente AU - Nagy, Ágota AU - Pál, Margit AU - Deák, Péter TI - Usp14 is required for spermatogenesis and ubiquitin stress responses in Drosophila melanogaster. JF - JOURNAL OF CELL SCIENCE J2 - J CELL SCI VL - 133 PY - 2020 IS - 2 PG - 6 SN - 0021-9533 DO - 10.1242/jcs.237511 UR - https://m2.mtmt.hu/api/publication/31158977 ID - 31158977 AB - Deubiquitylating (DUB) enzymes free covalently linked ubiquitin moieties from ubiquitin-ubiquitin and ubiquitin-protein conjugates, and thereby maintain the equilibrium between free and conjugated ubiquitin moieties and regulate ubiquitin-mediated cellular processes. Here, we performed genetic analyses of mutant phenotypes in Drosophila melanogaster and demonstrate that loss of Usp14 function results in male sterility, with defects in spermatid individualization and reduced testicular free monoubiquitin levels. These phenotypes were rescued by germline-specific overexpression of wild-type Usp14. Synergistic genetic interactions with Ubi-p63E and cycloheximide sensitivity suggest that ubiquitin shortage is a primary cause of male sterility. In addition, Usp14 is predominantly expressed in testes in Drosophila, indicating a higher demand for this DUB in testes that is also reflected by testis-specific loss-of-function Usp14 phenotypes. Collectively, these results suggest a major role of Usp14 in maintaining normal steady state free monoubiquitin levels during the later stages of Drosophila spermatogenesis.This article has an associated First Person interview with the first author of the paper. LA - English DB - MTMT ER -