FHL2 anchors mitochondria to actin and adapts mitochondrial dynamics to glucose supply

Basu, Himanish; Pekkurnaz, Gülçin; Falk, Jill; Wei, Wei; Chin, Morven; Steen, Judith A.; Schwarz, Thomas L.

doi:10.1083/jcb.201912077

Cited by 33 publications

(39 citation statements)

References 106 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…This perinuclear localization of peroxisomes was evident in transfections where the expressed mCitrine-KIF5C was not on peroxisomes (PEX3-C and TRAK1 or PEX3Miro1 without TRAK1). In contrast, when we expressed PEX3Miro1 with myc-TRAK1 and mCitrine-KIF5C, peroxisomes moved more (as in (Basu et al, 2021)) and were more widely dispersed in the cell (Figure 1E). We quantified their distribution using a lab-developed image-analysis FIJI macro, DoveSonoPro (Basu et al, 2021).…”

Section: Resultsmentioning

confidence: 96%

“…In contrast, when we expressed PEX3Miro1 with myc-TRAK1 and mCitrine-KIF5C, peroxisomes moved more (as in (Basu et al, 2021)) and were more widely dispersed in the cell (Figure 1E).…”

Section: Mislocalization Of Miro1 Is Sufficient For Co-localization O...mentioning

confidence: 94%

“…Constructs cloned in this study: 1. To make the PEX3-6xhis-mRFP construct (PEX3-C), the PEX3-6xhis-mRFP-Miro1 construct (PEX3Miro1) described in Basu et al, 2021 was linearized by PCR using Primers KP14 and KP15 to amplify the PEX3-6xhis-mRFP segment of the plasmid and the plasmid backbone but omit Miro1. The PCR product was ligated using the KLD reagent from the Q5 site-directed mutagenesis kit from NEB.…”

Section: Methodsmentioning

confidence: 99%

“…Quantifications of organelle distribution in COS7 cells: Quantification of peroxisome distribution was done as described in (Basu et al, 2021) using the DoveSonoPro software (https://github.com/ThomasSchwarzLab). DoveSonoPro software allows for the cell outline and cell center to be selected by the user.…”

Section: Methodsmentioning

confidence: 99%

“…We quantified their distribution using a lab-developed image-analysis FIJI macro, DoveSonoPro (Basu et al, 2021). DoveSonoPro measures the % frequency of objects as they appear in concentric shells moving out from the center of the cell.…”

Section: Mislocalization Of Miro1 Is Sufficient For Co-localization O...mentioning

confidence: 99%

See 4 more Smart Citations

Miro GTPase domains regulate assembly of the mitochondrial motor-adaptor complex

Davis

Basu

Shurberg

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

Mitochondrial transport relies on a motor-adaptor complex containing Miro1, a mitochondrial outer membrane protein with two GTPase domains, as well as TRAK1/2, kinesin-1, and dynein. Using a peroxisome-directed Miro1, we quantified the ability of GTPase mutations to influence peroxisomal recruitment of complex components. Miro1 whose N-GTPase is locked in the GDP-state doesn’t recruit TRAK1/2, kinesin or P135 to peroxisomes whereas the GTP-state does. Miro1 C-GTPase mutations have little influence on complex recruitment. Though Miro2 is thought to support mitochondrial motility, peroxisome-directed Miro2 did not recruit the other complex components regardless of the state of its GTPase domains. Neurons expressing peroxisomal Miro1 with the GTP-state form of the N-GTPase had markedly increased peroxisomal transport to growth cones while the GDP-state caused their retention in the soma. Thus, the N-GTPase of Miro1 is critical for regulating Miro1’s interaction with the other components of the motor-adaptor complex and thereby for regulating mitochondrial motility.SummaryA Miro-containing complex mediates mitochondrial motility. Relocalizing Miro1 and 2 to peroxisomes and systematically manipulating each GTPase domain of Miro revealed the importance of the N-terminal GTPase domain of Miro1 for governing interaction with TRAK proteins, motors, and transport.

show abstract

Section: Resultsmentioning

confidence: 96%

“…In contrast, when we expressed PEX3Miro1 with myc-TRAK1 and mCitrine-KIF5C, peroxisomes moved more (as in (Basu et al, 2021)) and were more widely dispersed in the cell (Figure 1E).…”

Section: Mislocalization Of Miro1 Is Sufficient For Co-localization O...mentioning

confidence: 94%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Mislocalization Of Miro1 Is Sufficient For Co-localization O...mentioning

confidence: 99%

See 3 more Smart Citations

Miro GTPase domains regulate assembly of the mitochondrial motor-adaptor complex

Davis

Basu

Shurberg

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

Protease-independent control of parthanatos by HtrA2/Omi

Weiß,

Heib,

Korn

et al. 2023

Cell. Mol. Life Sci.

View full text Add to dashboard Cite

HtrA2/Omi is a mitochondrial serine protease with ascribed pro-apoptotic as well as pro-necroptotic functions. Here, we establish that HtrA2/Omi also controls parthanatos, a third modality of regulated cell death. Deletion of HtrA2/Omi protects cells from parthanatos while reconstitution with the protease restores the parthanatic death response. The effects of HtrA2/Omi on parthanatos are specific and cannot be recapitulated by manipulating other mitochondrial proteases such as PARL, LONP1 or PMPCA. HtrA2/Omi controls parthanatos in a manner mechanistically distinct from its action in apoptosis or necroptosis, i.e., not by cleaving cytosolic IAP proteins but rather exerting its effects without exiting mitochondria, and downstream of PARP-1, the first component of the parthanatic signaling cascade. Also, previously identified or candidate substrates of HtrA2/Omi such as PDXDC1, VPS4B or moesin are not cleaved and dispensable for parthanatos, whereas DBC-1 and stathmin are cleaved, and thus represent potential parthanatic downstream mediators of HtrA2/Omi. Moreover, mass-spectrometric screening for novel parthanatic substrates of HtrA2/Omi revealed that the induction of parthanatos does not cause a substantial proteolytic cleavage or major alterations in the abundance of mitochondrial proteins. Resolving these findings, reconstitution of HtrA2/Omi-deficient cells with a catalytically inactive HtrA2/Omi mutant restored their sensitivity against parthanatos to the same level as the protease-active HtrA2/Omi protein. Additionally, an inhibitor of HtrA2/Omi’s protease activity did not confer protection against parthanatic cell death. Our results demonstrate that HtrA2/Omi controls parthanatos in a protease-independent manner, likely via novel, unanticipated functions as a scaffolding protein and an interaction with so far unknown mitochondrial proteins.

show abstract