Structural studies of human fission protein FIS1 reveal a dynamic region important for GTPase DRP1 recruitment and mitochondrial fission

Egner, John M.; Nolden, Kelsey A; Harwig, Megan Cleland; Bonate, Ryan P.; Anda, Jaime de; Tessmer, Maxx H.; Noey, Elizabeth L.; Ihenacho, Ugochukwu K.; Liu, Ziwen; Peterson, Francis C.; Wong, Gerard C. L.; Widlansky, Michael E.; Hill, Rolla B.

doi:10.1016/j.jbc.2022.102620

Cited by 11 publications

(41 citation statements)

References 84 publications

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“…To evaluate this, we immunostained these cells for DRP1 and quantified colocalization with the MitoYFP signal ( Figure 3C ). Mitochondrial recruitment of DRP1 is potently induced upon ectopic FIS1 overexpression consistent with earlier findings (15, 16, 41) ( Figure 3D ). Therefore, we asked if mitochondrial DRP1 recruitment was significantly perturbed between wild-type and variant FIS1 overexpression.…”

Section: Resultssupporting

confidence: 92%

“…Batch mode pre-processing of images for mitochondrial area assessment by MitoGraph was done using R scripts previously described (41, 59). MitoGraph segmentation and noise removal were performed on cropped TIFF files using the following commands for segmentation: MitoGraph -xy 0.07 -z 0.2 -adaptive 10 -path cells.…”

Section: Methodsmentioning

confidence: 99%

“…MitoGraph segmentation and noise removal were performed on cropped TIFF files using the following commands for segmentation: MitoGraph -xy 0.07 -z 0.2 -adaptive 10 -path cells. The resulting PNG files were compiled using an ImageJ macro and screened for accurate mitochondrial segmentation as previously described (41). The average mitochondrial area was then determined by multiplying the average edge length and average width values generated by MitoGraph.…”

Section: Image Acquisition Colocalization Lut Intensity and Mitochond...mentioning

confidence: 99%

“…FIS1 is an atypical TPR protein because it possesses two repeats, only one of which is canonical (35,36). Furthermore, FIS1 exists in oligomeric heterocomplexes mediated by its TPRs, which may be auto-inhibited by its N-terminal helix as deletion of this helix enhances FIS1 oligomerization and DRP1 recruitment (17,23,(39)(40)(41). Adjacent to the N-terminal helix is a disordered region of FIS1, termed the FIS1 arm, that is required for its mitochondrial fission functions in both yeast and human cells (41,42).…”

Section: Introductionmentioning

confidence: 99%

“…Furthermore, FIS1 exists in oligomeric heterocomplexes mediated by its TPRs, which may be auto-inhibited by its N-terminal helix as deletion of this helix enhances FIS1 oligomerization and DRP1 recruitment (17,23,(39)(40)(41). Adjacent to the N-terminal helix is a disordered region of FIS1, termed the FIS1 arm, that is required for its mitochondrial fission functions in both yeast and human cells (41,42). Consistent with a key role for the N-terminal region are splice variants in mice and fruitflies that lack this region (43).…”

Section: Introductionmentioning

confidence: 99%

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A conserved, non-canonical insert in FIS1 is required for mitochondrial fission and recruitment of DRP1 and TBC1D15

Ihenacho

Toro

Mansour

et al. 2023

Preprint

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Mitochondrial fission protein 1 (FIS1) is conserved in all eukaryotes yet its activity in metazoans is thought divergent from lower eukaryotes like fungi. To address this discrepancy, structure-based sequence alignments revealed a conserved but non-canonical, three-residue insert in a FIS1 turn suggesting a conserved activity. In vertebrate FIS1 this insert is serine (S45), lysine (K46), and tyrosine (Y47). To determine the biological role of this SKY insert, three variants were evaluated for their fold, and tested in HCT116 cells for altered mitochondrial morphology and recruitment of effectors, DRP1 and TBC1D15. Substitution of the SKY insert with three alanine residues (AAA), or deletion of the insert (ΔSKY), did not substantially alter the fold or thermal stability of the protein. Replacing SKY with a canonical turn (ΔSKYD49G) introduced significant conformational heterogeneity by NMR that was removed upon deletion of a known regulatory region, the Fis1 arm. Expression of AAA fragmented mitochondria into perinuclear clumps associated with increased mitochondrial Drp1 similar to the wild-type protein. In contrast, expression of ΔSKY variants elongated mitochondrial networks and reduced mitochondrial Drp1. Co-expression of YFP-TBC1D15 partially rescued mitochondrial morphology and Drp1 recruitment for ΔSKY variants, although ΔSKY variants were markedly unable to support TBC1D15 assembly into punctate structures found upon co-expression with wildtype Fis1 or the AAA variant. Collectively these results show that FIS1 activity can be modulated by conserved residues supporting a generalized model whereby FIS1 is governed by intramolecular interactions between the regulatory FIS1 arm and SKY insert that may be conserved across species.

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Section: Resultssupporting

confidence: 92%

Section: Methodsmentioning

confidence: 99%

Section: Image Acquisition Colocalization Lut Intensity and Mitochond...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A conserved, non-canonical insert in FIS1 is required for mitochondrial fission and recruitment of DRP1 and TBC1D15

Ihenacho

Toro

Mansour

et al. 2023

Preprint

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Asiaticoside alleviates lipopolysaccharide-induced acute lung injury by blocking Sema4D/CD72 and inhibiting mitochondrial dysfunction in RAW264.7 cell and mice

Zhang,

Zhao,

Wang

et al. 2024

Naunyn-Schmiedeberg's Arch Pharmacol

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Acute lung injury (ALI) is a common disease with complex pathogenesis. However, the treatment is mainly symptomatic with limited clinical options. Asiaticoside (AS), a Chinese herbal extract, has protective effects against LPS-induced ALI in mice and inhibits nitric oxide and prostaglandin E2 synthesis; however, the specific mechanism of AS in the prevention and treatment of LPS-induced ALI needs further study. Sema4D/CD72 pathway, mitochondrial dysfunction, and miRNA-21 are closely associated with inflammation. Therefore, the present study aimed to explore whether AS exerts its therapeutic effect on ALI by influencing Sema4D/CD72 pathway and mitochondrial dysfunction, restoring the balance of inflammatory factors, and influencing miRNA-21 expression. Cell and animal experiments were performed to investigate the effect of AS on ALI. Lipopolysaccharide (LPS) was used to establish the ALI model. CCK8 and flow cytometry were used to detect the cell viability and apoptosis rate. HE staining and wet-to-dry weight ratio (W/D) of lung tissue were determined. The expressions of Sema4D, CD72, NF-κB p65, Bax, Bcl2, and caspase 3 in RAW264.7 cells and lung tissues were detected by western blot, and the levels of IL-10 and IL-1β induced by LPS in supernatant of RAW264.7 cells and BALF were measured by ELISA. And the expression of miRNA-21 in cells and lung tissues was detected by fluorescence quantitative PCR. The result shows that AS treatment suppressed LPS-induced cell damage and lung injury in mice. AS treatment could alleviate the pathological changes such as inflammatory infiltration and histopathological changes in the lungs caused by LPS, and reduce the ratio of W/D. AS significantly alleviated the decrease of mitochondrial membrane potential induced by LPS, inhibited the increase of ROS production, and reduced the expression of mitochondrial fission proteins Drp1 and Fis1. The high-dose AS group significantly downregulated the expression of Sema4D, CD72, phosphorylated NF-κB p65, and apoptosis-related proteins, decreased the pro-inflammatory factor IL-1β, and enhanced the level of anti-inflammatory factor IL-10. In addition, AS promoted miRNA-21 expression. These effects inhibited apoptosis and restored the balance between anti- and pro-inflammatory factors. This represents the inaugural report elucidating the mechanism by which AS inhibits the Sema4D/CD72 signaling pathway. These findings offer novel insights into the potential application of AS in both preventing and treating ALI.

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A conserved, noncanonical insert in FIS1 mediates TBC1D15 and DRP1 recruitment for mitochondrial fission

Ihenacho,

Toro,

Mansour

et al. 2023

Journal of Biological Chemistry

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Structural studies of human fission protein FIS1 reveal a dynamic region important for GTPase DRP1 recruitment and mitochondrial fission

Cited by 11 publications

References 84 publications

A conserved, non-canonical insert in FIS1 is required for mitochondrial fission and recruitment of DRP1 and TBC1D15

A conserved, non-canonical insert in FIS1 is required for mitochondrial fission and recruitment of DRP1 and TBC1D15

Asiaticoside alleviates lipopolysaccharide-induced acute lung injury by blocking Sema4D/CD72 and inhibiting mitochondrial dysfunction in RAW264.7 cell and mice

A conserved, noncanonical insert in FIS1 mediates TBC1D15 and DRP1 recruitment for mitochondrial fission

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