Rab40c regulates focal adhesions and PP6 activity by controlling ANKRD28 ubiquitylation

Han, Ke-Jun; Mikalayeva, Valeryia; Gerber, Scott A.; Kettenbach, Arminja N.; Skeberdis, Vytenis Arvydas; Prekeris, Rytis

doi:10.26508/lsa.202101346

Cited by 7 publications

(8 citation statements)

References 64 publications

(96 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

Section: Discussionmentioning

confidence: 99%

“…An emerging function of PP6 is as a negative regulator of the Hippo signaling pathway (39, 40, 52). PP6 subunits interact with MOB1 in a phosphorylation-dependent manner (39, 40), and long-term depletion of ANKRD28 increased MOB1 T35 phosphorylation, regulating focal adhesion dynamics in cell migration (52). Our analysis revealed a significant increase in S9 and T35 MOB1 phosphorylation upon PP6c degradation, as well as an enrichment in the cellular component category for cell junctions.…”

Section: Discussionmentioning

confidence: 99%

“…The PP6 holoenzyme is comprised of a catalytic subunit in a complex with one of three Sit4-associated protein (SAP) subunits, SAPS1, 2, or 3 (also referred to as PP6R1-3), as well as with one of three Ankyrindomain containing proteins, ANKRD28, 44 or 52 (15,16). PP6 holoenzymes have been implicated in chromosome condensation and segregation in mitosis (17)(18)(19)(20)(21)(22)(23), cell cycle progression (24), inflammatory response (25)(26)(27)(28), DNA damage (29-35), splicing (36), maintenance of cell contacts (37,38), and Hippo (38)(39)(40) and mitogen-activated kinase (MAPK) signaling (41). In cancer, PP6C is mutated and deregulated in melanoma (42)(43)(44)(45)(46)(47).…”

Section: Introductionmentioning

confidence: 99%

“…MST1/2 bind MOB1 in a phosphorylation-dependent manner and phosphorylate MOB1 on Threonine 12 (T12) and Threonine 35 (T35), inducing a conformational shift that exposes a binding surface for LATS and NDR kinases (51). Long-term depletion of ANKRD28 by siRNA increased MOB1 T35 phosphorylation (52). However, a direct regulatory function of PP6 holoenzymes in the Hippo pathway has not been demonstrated.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Inducible protein degradation as a strategy to identify Phosphoprotein Phosphatase 6 substrates in RAS-mutant colorectal cancer cells

Mariano

Rusin

Nasa

et al. 2023

Preprint

View full text Add to dashboard Cite

Protein phosphorylation is an essential regulatory mechanism that controls most cellular processes, including cell cycle progression, cell division, and response to extracellular stimuli, among many others, and is deregulated in many diseases. Protein phosphorylation is coordinated by the opposing activities of protein kinases and protein phosphatases. In eukaryotic cells, most serine/threonine phosphorylation sites are dephosphorylated by members of the Phosphoprotein Phosphatase (PPP) family. However, we only know for a few phosphorylation sites which specific PPP dephosphorylates them. Although natural compounds such as calyculin A and okadaic acid inhibit PPPs at low nanomolar concentrations, no selective chemical PPP inhibitors exist. Here, we demonstrate the utility of endogenous tagging of genomic loci with an auxin-inducible degron (AID) as a strategy to investigate specific PPP signaling. Using Protein Phosphatase 6 (PP6) as an example, we demonstrate how rapidly inducible protein degradation can be employed to identify dephosphorylation SITES and elucidate PP6 biology. Using genome editing, we introduce AID-tags into each allele of the PP6 catalytic subunit (PP6c) in DLD-1 cells expressing the auxin receptor Tir1. Upon rapid auxin-induced degradation of PP6c, we perform quantitative mass spectrometry-based proteomics and phosphoproteomics to identify PP6 substrates in mitosis. PP6 is an essential enzyme with conserved roles in mitosis and growth signaling. Consistently, we identify candidate PP6c-dependent phosphorylation sites on proteins implicated in coordinating the mitotic cell cycle, cytoskeleton, gene expression, and mitogen-activated protein kinase (MAPK) and Hippo signaling. Finally, we demonstrate that PP6c opposes the activation of large tumor suppressor 1 (LATS1) by dephosphorylating Threonine 35 (T35) on Mps One Binder (MOB1), thereby blocking the interaction of MOB1 and LATS1. Our analyses highlight the utility of combining genome engineering, inducible degradation, and multiplexed phosphoproteomics to investigate signaling by individual PPPs on a global level, which is currently limited by the lack of tools for specific interrogation.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Inducible protein degradation as a strategy to identify Phosphoprotein Phosphatase 6 substrates in RAS-mutant colorectal cancer cells

Mariano

Rusin

Nasa

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…Moreover, the same authors found RAB40B to be highly expressed in more aggressive cancers, as well as in basal BC subtype. Additionally, RAB40C was found to regulate focal adhesion number, size, and distribution in migrating BC cells ( Han et al, 2022 ).…”

Section: Secretion/exocytosismentioning

confidence: 99%

Membrane trafficking alterations in breast cancer progression

Ferreira,

Castanheira,

Escrevente

et al. 2024

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

Breast cancer (BC) is the most common type of cancer in women, and remains one of the major causes of death in women worldwide. It is now well established that alterations in membrane trafficking are implicated in BC progression. Indeed, membrane trafficking pathways regulate BC cell proliferation, migration, invasion, and metastasis. The 22 members of the ADP-ribosylation factor (ARF) and the >60 members of the rat sarcoma (RAS)-related in brain (RAB) families of small GTP-binding proteins (GTPases), which belong to the RAS superfamily, are master regulators of membrane trafficking pathways. ARF-like (ARL) subfamily members are involved in various processes, including vesicle budding and cargo selection. Moreover, ARFs regulate cytoskeleton organization and signal transduction. RABs are key regulators of all steps of membrane trafficking. Interestingly, the activity and/or expression of some of these proteins is found dysregulated in BC. Here, we review how the processes regulated by ARFs and RABs are subverted in BC, including secretion/exocytosis, endocytosis/recycling, autophagy/lysosome trafficking, cytoskeleton dynamics, integrin-mediated signaling, among others. Thus, we provide a comprehensive overview of the roles played by ARF and RAB family members, as well as their regulators in BC progression, aiming to lay the foundation for future research in this field. This research should focus on further dissecting the molecular mechanisms regulated by ARFs and RABs that are subverted in BC, and exploring their use as therapeutic targets or prognostic markers.

show abstract