Biology subjects

Ziv, T.

Publications and source records attributed to Ziv, T..

5 recordsLinked to original sources

bioRxiv2025

Succinate and its carrier Sfc1 mediate metabolic control of mitochondrial protein import by the TIM23 translocase

Tim23 is an essential component of the mitochondrial inner membrane translocase and Sfc1 is a carrier that exchanges succinate for fumarate across that membrane. Sfc1 and succinic acid availability regulate dual targeting of fumarase and aconitase by facilitating mitochondrial import of their newly synthesized precursors, as shown by pulse-chase experiments. Here we show that Sfc1 directly interacts with the Tim23 and succinate affects this interaction, which in turn affects mitochondrial protein import. Physical interaction between Tim23 and Sfc1 was proven by co immunoprecipitation, Bimolecular Fluorescence Complementation (BiFC) and Biotin-based proximity labeling (TurboID). Proximity labeling-informed mutagenesis allowed us to dissect the carrier activity of Sfc1 from its function as a TIM23 regulator. We performed Rosetta-MP docking of Sfc1 and Tim23 to envisage the interface. Thus, our findings show that metabolites can regulate mitochondrial import and adjust the segregation of key metabolic enzymes between the cytosol and mitochondria.

cell biology↗

bioRxiv2025

When Less is MORE: Counterintuitive Stoichiometries and Cellular Abundances are Essential for ABC transporters function

Prokaryotes acquire essential nutrients primarily through ABC importers, consisting of an ATPase, a permease, and a substrate-binding protein. These importers are highly underrepresented in proteomic databases, limiting our knowledge about their cellular copy numbers, component stoichiometry, and the mechanistic implications of these parameters. We developed a tailored proteomic approach to compile the most comprehensive dataset to date of the E. coli ABC importome. Functional assays and analysis of deletion strains revealed novel mechanistic features, linking molecular mechanisms to cellular abundances, co-localization, and component stoichiometries. We observed 4-5 orders of magnitude variation in import system abundances, with copy numbers tuned to nutrient hierarchies essential for growth. Abundances of substrate-binding proteins are unrelated to their substrate binding affinities but are tightly, yet inversely, correlated with their interaction affinity with permeases. Counterintuitive component stoichiometries are crucial for function, offering insights into the design principles of multi-component protein systems, potentially extending beyond ABC importers. TeaserBacterias nutrient absorption secrets unveiled: deciphering the complexity of ABC transporter systems required for optimal growth.

biochemistry↗

bioRxiv2023

Ribosomal Protein SA (RPSA) is required for localized translation and sarcomere maintenance

Cardiomyocyte sarcomeres contain localized ribosomes, but the factors responsible for their localization and the significance of localized translation are unknown. Using proximity labeling, we identified Ribosomal Protein SA (RPSA) as a Z-line protein. In cultured cardiomyocytes, the loss of RPSA led to impaired local protein translation and reduced sarcomere integrity. By employing CAS9 expressing mice along with adeno-associated viruses expressing CRE recombinase and single-guide RNAs targeting Rpsa, we knocked out Rpsa in vivo and observed mis-localization of ribosomes and diminished local translation. These genetic mosaic mice with Rpsa knockout in a subset of cardiomyocytes developed dilated cardiomyopathy, featuring atrophy of RPSA-deficient cardiomyocytes, compensatory hypertrophy of unaffected cardiomyocytes, left ventricular dilation, and impaired contractile function. We demonstrate that RPSA C-terminal domain is sufficient for localization to the Z-lines and that if the microtubule network is disrupted RPSA loses its sarcomeric localization. These findings highlight RPSA as a ribosomal factor essential for ribosome localization to the Z-line, facilitating local translation and sarcomere maintenance.

cell biology↗

bioRxiv2022

Yeast Mitochondrial Derived Vesicles (MDVs) have membrane potential and contain a functional ATP synthase

Vesicular transport is a means of communication. While cells can communicate between each other via secretion of extracellular vesicles, less is known regarding organelle-to organelle communication, in particularly in the case of mitochondria. Mitochondria are responsible for the production of energy and for essential metabolic pathways in the cell, as well as fundamental processes such as apoptosis and aging. Here we show that functional mitochondria, isolated from Saccharomyces cerevisiae release vesicles, independent of the fission machinery. We were then able for the first time to isolate these Mitochondrial Derived Vesicles (MDVs) and found that they are relatively uniform in size, of about 100nm and carry selective protein cargo including enrichment of ATP synthase subunits. Remarkably, we further found that these MDVs harbor a functional ATP synthase complex. Moreover, we demonstrate that these vesicles have a membrane potential, produce ATP, and seem to fuse with naive mitochondria. Our findings reveal a possible delivery mechanism of ATP producing vesicles, which can potentially regenerate ATP deficient mitochondria and may participate in organelle to organelle communication.

cell biology↗

bioRxiv2019

Numerous post-translational modifications of RNA polymerase II subunit Rpb4 link transcription to post-transcriptional mechanisms

Rpb4/7 binds RNA Polymerase II (Pol II) transcripts co-transcriptionally and accompanies them throughout their lives. By virtue of its capacity to interact with key regulators (e.g., Pol II, eIF3, Pat1) both temporarily and spatially, Rpb4/7 regulates the major stages of the mRNA lifecycle. Here we show that Rpb4/7 can undergo over 100 combinations of post-translational modifications (PTMs). Remarkably, the Rpb4/7 PTMs repertoire changes as the mRNA/Rpb4/7 complex progresses from one stage to the next. A mutagenesis approach in residues that undergo PTMs suggests that temporal Rpb4 PTMs regulate its interactions with key regulators of gene expression that control transcriptional and post-transcriptional stages. Moreover, one mutant type specifically affects mRNA synthesis despite its normal association with Pol II, whereas the other affects both mRNA synthesis and decay; both types disrupt the balance between mRNA synthesis and decay ( mRNA buffering) and the cells capacity to respond to the environment. Taken together, we propose that temporal Rpb4/7 PTMs are involved in cross talks among the various stages of the mRNA lifecycle.

molecular biology↗