Structural Basis for Rab8a Recruitment of RILPL2 via LRRK2 Phosphorylation of Switch 2

Waschbüsch, Dieter; Purlyte, Elena; Pal, Prosenjit; McGrath, Emma; Alessi, Dario R.; Khan, Amir R.

doi:10.1016/j.str.2020.01.005

Cited by 71 publications

(122 citation statements)

References 48 publications

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“…Like RILPL2, the related RILPL1 also binds preferentially to phosphorylated Rab8 and Rab10 (Steger et al, 2017;Waschbüsch et al, 2020). We showed previously that RILPL1 enhances the pericentriolar clustering of pRab10-containing membrane compartments .…”

Section: Myova Colocalizes With Prab10 Independent Of Rilpl2mentioning

confidence: 85%

“…Bacterially expressed, purified Rab10 was phosphorylated in vitro using Mst3, a promiscuous kinase used previously to phosphorylate purified Rab8 and Rab10 proteins (Berndsen et al, 2019;Waschbüsch et al, 2020). In vitro phosphorylation of Rab10 proceeded linearly for ~100 minutes before reaching a plateau in reactions containing 2mM ATP and a ratio of 1:3 kinase:substrate at 27°C, as detected using a phosphoRab10-specific antibody (Fig.…”

Section: Purified Myova Globular Tail Domain Binds Phosphorylated Rabmentioning

confidence: 99%

“…RILPL2 interacts with the Cterminal, globular tail domain of MyoVa via its N-terminal "RH1" domain (Lisé et al, 2009) and binds pRab8 and pRab10 (Steger et al, 2017) via its C-terminal RH2 domain (129-165; cf. Waschbüsch et al, 2020). Recently, RILPL2 has also been reported to regulate the motor activity of MyoVa (Cao et al, 2019).…”

Section: Phosphorylation Of Rab Proteins Blocks Their Abilities To Inmentioning

confidence: 99%

“…These data are consistent with pRab10 binding both RILPL2 and MyoVa, perhaps forming a ternary complex. Since RILPL2 can dimerize (Wei et al, 2013;Waschbüsch et al, 2020), it is possible that each half of the dimer engages either MyoVa or pRab10 or both. Alternatively, pRab10 may enhance the ability of MyoVa to bind RILPL2.…”

Section: Prab10 Binding To the Myova Globular Tail Domainmentioning

confidence: 99%

“…Despite its ability to bind RILPL2, endogenous Rab8 which is also phosphorylated by R1441G LRRK2 and located at the centriole (Purlyte et al, 2018;Ordonez et al, 2019) does not appear to be sufficient to drive RILPL2 relocalization in cells depleted of Rab10. This may be because MyoVa binds with preference to Rab10, and the MyoVa globular tail domain binds with strong preference to pRab10 and apparently not to pRab8 (Waschbüsch et al, 2020).…”

Section: Rilpl2 Binds Lrrk2-phosphorylated Rab10 With Greater Affinitmentioning

confidence: 99%

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LRRK2-phosphorylated Rab10 sequesters Myosin Va with RILPL2 during ciliogenesis blockade

Izumi

Dhekne

Vides

et al. 2020

Preprint

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Activating mutations in LRRK2 kinase cause Parkinson's disease. Pathogenic LRRK2 phosphorylates a subset of Rab GTPases and blocks ciliogenesis. Thus, defining novel phospho-Rab interacting partners is critical to our understanding of the molecular basis of LRRK2 pathogenesis. RILPL2 binds with strong preference to LRRK2-phosphorylated Rab8A and Rab10. RILPL2 is a binding partner of the motor protein and Rab effector, Myosin Va. We show here that the globular tail domain of Myosin Va also contains a high affinity binding site for LRRK2-phosphorylated Rab10, and certain tissue-specific Myosin Va isoforms strongly prefer to bind phosphorylated Rab10. In the presence of pathogenic LRRK2, RILPL2 relocalizes to the peri-centriolar region in a phosphoRab10-and Myosin Vadependent manner. In the absence of phosphoRab10, expression of RILPL2 or depletion of Myosin Va increase centriolar RILPL2 levels, and either condition is sufficient to block ciliogenesis in RPE cells. These experiments show that LRRK2 generated phosphoRab10 dramatically redistributes Myosin Va-RILPL2 complexes to the mother centriole, which may sequester Myosin Va and RILPL2 in a manner that blocks their normal roles in ciliogenesis.

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Section: Myova Colocalizes With Prab10 Independent Of Rilpl2mentioning

confidence: 85%

Section: Purified Myova Globular Tail Domain Binds Phosphorylated Rabmentioning

confidence: 99%

Section: Phosphorylation Of Rab Proteins Blocks Their Abilities To Inmentioning

confidence: 99%

Section: Prab10 Binding To the Myova Globular Tail Domainmentioning

confidence: 99%

Section: Rilpl2 Binds Lrrk2-phosphorylated Rab10 With Greater Affinitmentioning

confidence: 99%

See 3 more Smart Citations

LRRK2-phosphorylated Rab10 sequesters Myosin Va with RILPL2 during ciliogenesis blockade

Izumi

Dhekne

Vides

et al. 2020

Preprint

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LRRK2 phosphorylation of Rab GTPases in Parkinson's disease

Pfeffer

2022

FEBS Letters

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Rab GTPases comprise a large family of conserved GTPases that are critical regulators of the secretory and endocytic pathways. The human genome encodes ~ 65 Rabs that localize to discrete membrane compartments and, when in their GTP‐bound state, bind to effector proteins to carry out diverse functions. Activating mutations in LRRK2 kinase cause Parkinson's disease, and subsets of Rab GTPases are important LRRK2 substrates. LRRK2 phosphorylates a conserved threonine residue that is essential for Rab interaction with guanine nucleotide exchange factors, effectors, and GDI that recycles Rabs between membrane compartments. This brief review will highlight new findings related to LRRK2‐mediated phosphorylation of Rab GTPases and its consequences. Remarkably, Rab phosphorylation flips a switch on Rab effector selection with dominant consequences for cell pathophysiology.

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Targeting Rab‐RILPL interactions as a strategy to downregulate pathogenic LRRK2 in Parkinson's disease

Alexander,

Naaldijk,

Fasiczka

et al. 2023

Journal of Peptide Science

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Familial Parkinson's disease (PD) is frequently linked to multiple disease‐causing mutations within Leucine‐Rich Repeat Protein Kinase 2 (LRRK2), leading to aberrant kinase activity. Multiple pathogenic effects of enhanced LRRK2 activity have been identified, including loss of cilia and centrosomal cohesion defects. When phosphorylated by LRRK2, Rab8a and Rab10 bind to phospho‐specific RILPL effector proteins. RILPL‐mediated accumulation of pRabs proximal to the mother centriole is critical for initiating deficits in ciliogenesis and centrosome cohesion mediated by LRRK2. We hypothesized that Rab‐derived phospho‐mimics may serve to block phosphorylated Rab proteins from docking with RILPL in the context of hyperactive LRRK2 mutants. This would serve as an alternative strategy to downregulate pathogenic signaling mediated by LRRK2, rather than targeting LRRK2 kinase activity itself. To test this theory, we designed a series of constrained peptides mimicking phosphorylated Switch II derived from Rab8. These RILPL interacting peptides, termed RIP, were further shown to permeate cells. Further, several peptides were found to bind RILPL2 and restore ciliogenesis and centrosomal cohesion defects in cells expressing PD‐associated mutant LRRK2. This research demonstrates the utility of constrained peptides as downstream inhibitors to target pathogenic LRRK2 activity and may provide an alternative approach to target specific pathways activated by LRRK2.

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Structural Basis for Rab8a Recruitment of RILPL2 via LRRK2 Phosphorylation of Switch 2

Cited by 71 publications

References 48 publications

LRRK2-phosphorylated Rab10 sequesters Myosin Va with RILPL2 during ciliogenesis blockade

LRRK2-phosphorylated Rab10 sequesters Myosin Va with RILPL2 during ciliogenesis blockade

LRRK2 phosphorylation of Rab GTPases in Parkinson's disease

Targeting Rab‐RILPL interactions as a strategy to downregulate pathogenic LRRK2 in Parkinson's disease

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