Endosomal-associated RFFL facilitates mitochondrial clearance by enhancing PRKN/parkin recruitment to mitochondria

Ravindran, Rishith; Velikkakath, Anoop Kumar G.; Narendradev, Nikhil Dev; Chandrasekharan, Aneesh; Santhoshkumar, T.R.; Srinivasula, Srinivasa M.

doi:10.1080/15548627.2022.2052460

Cited by 3 publications

(2 citation statements)

References 57 publications

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“…HeLa cells obtained from American Type Culture Collection (ATCC) were cultured and stained with mitotracker as described in our previously reported study 43 . To mount the sample vertically to be adaptable in our home-built (M -sMx-SPIM) microscopy system, the cell was cultured in a glass bottom dish (Cell Vis, D35-20-1.5-N).…”

Section: Methodsmentioning

confidence: 99%

Real-time study of spatio-temporal dynamics (4D) of physiological activities in alive biological specimens with different FOVs and resolutions simultaneously

K. S.,

Sarkar,

Vishnu

et al. 2024

Sci Rep

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This article reports the development of a microscopy imaging system that gives feasibility for studying spatio-temporal dynamics of physiological activities of alive biological specimens (over entire volume not only for a particular section, i.e., in 4D). The imaging technology facilitates to obtain two image frames of a section of the larger specimen ($$\sim \text {mm}$$ ∼ mm ) with different FOVs at different resolutions or magnifications simultaneously in real-time (in addition to recovery of 3D (volume) information). Again, this imaging system addresses the longstanding challenges of housing multiple light sources (6 at the maximum till date) in microscopy (in general) and light sheet fluorescence microscopy (LSFM) (in particular), by using a tuneable pulsed laser source (with an operating wavelength in the range $$\sim 420$$ ∼ 420 –670 nm) in contrast to the conventional CW laser source being adopted for inducing photo-excitation of tagged fluorophores. In the present study, we employ four wavelengths ($$\sim$$ ∼ 488 nm, 585 nm, 590 nm, and 594 nm). Our study also demonstrates quantitative characterization of spatio-temporal dynamics (velocity—both amplitude and direction) of organelles (mitochondria) and their mutual correlationships. Mitochondria close to the nucleus (or in clustered cells) are observed to possess a lower degree of freedom in comparison to that at the cellular periphery (or isolated cells). In addition, the study demonstrates real-time observation and recording of the development and growth of all tracheal branches during the entire period ($$\sim 95$$ ∼ 95 min) of embryonic development (Drosophila). The experimental results—with experiments being conducted in various and diversified biological specimens (Drosophila melanogaster, mouse embryo, and HeLa cells)—demonstrate that the study is of great scientific impact both from the aspects of technology and biological sciences.

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Section: Methodsmentioning

confidence: 99%

Real-time study of spatio-temporal dynamics (4D) of physiological activities in alive biological specimens with different FOVs and resolutions simultaneously

K. S.,

Sarkar,

Vishnu

et al. 2024

Sci Rep

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“…CARP2 is shown to be a negative regulator of NF‐kB and controls inflammation by targeting RIP kinase and is involved in the removal of damaged mitochondria, etc. [20,21]. We showed that CARPs interact and ubiquitinate STUB1, affecting STUB1 localization.…”

Section: Introductionmentioning

confidence: 95%

CARPs regulate STUB1 and its pathogenic mutants aggregation kinetics by mono‐ubiquitination

2023

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The development of neurological pathologies is linked to the accumulation of protein aggregates like alpha‐synuclein in Parkinson's disease and tau protein in Alzheimer's disease. Mono‐ or di‐ubiquitination of these molecules has been reported to stabilize aggregates and contribute to the disorders. STIP1 Homologous and U‐Box‐containing protein 1 (STUB1) is a multifunctional protein that maintains proteostasis and insulin signalling. In spinocerebellar ataxia 16 (SCAR16), an autosomal recessive neurodegenerative disease, mutations in and aggregation of STUB1 are reported. Despite the well‐accepted neuroprotective role of STUB1, very little is known of regulatory mechanisms that control the dynamics of STUB1 aggregate assembly. Here, we report that CARP2, a ubiquitin ligase, is a novel regulator of STUB1. CARP2 interacts and mono‐ubiquitinates STUB1. Furthermore, we found that CARP2 regulates STUB1 through its TPR motif, a domain that is also associated with HSP70. Modification of STUB1 by CARP2 leads to detergent‐insoluble aggregate formation. Importantly, pathogenic mutants of STUB1 are more prone than the wild‐type to CARP2‐mediated aggregate assembly. Hence our findings revealed CARPs (CARP1 & CARP2) as novel regulators of STUB1 and controlled its cytosolic versus aggregate dynamics.

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MCSdb, a database of proteins residing in membrane contact sites

Pan,

Ren,

Yang

et al. 2024

Sci Data

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Organelles do not act as autonomous discrete units but rather as interconnected hubs that engage in extensive communication by forming close contacts called “membrane contact sites (MCSs)”. And many proteins have been identified as residing in MCS and playing important roles in maintaining and fulfilling specific functions within these microdomains. However, a comprehensive compilation of these MCS proteins is still lacking. Therefore, we developed MCSdb, a manually curated resource of MCS proteins and complexes from publications. MCSdb documents 7010 MCS protein entries and 263 complexes, involving 24 organelles and 44 MCSs across 11 species. Additionally, MCSdb orchestrates all data into different categories with multitudinous information for presenting MCS proteins. In summary, MCSdb provides a valuable resource for accelerating MCS functional interpretation and interorganelle communication deciphering.

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Endosomal-associated RFFL facilitates mitochondrial clearance by enhancing PRKN/parkin recruitment to mitochondria

Cited by 3 publications

References 57 publications

Real-time study of spatio-temporal dynamics (4D) of physiological activities in alive biological specimens with different FOVs and resolutions simultaneously

Real-time study of spatio-temporal dynamics (4D) of physiological activities in alive biological specimens with different FOVs and resolutions simultaneously

CARPs regulate STUB1 and its pathogenic mutants aggregation kinetics by mono‐ubiquitination

MCSdb, a database of proteins residing in membrane contact sites

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