scholarly journals Membrane association but not identity is required for LRRK2 activation and phosphorylation of Rab GTPases

2019 ◽  
Vol 218 (12) ◽  
pp. 4157-4170 ◽  
Author(s):  
Rachel C. Gomez ◽  
Paulina Wawro ◽  
Pawel Lis ◽  
Dario R. Alessi ◽  
Suzanne R. Pfeffer
Keyword(s):  
Plasma Membrane ◽  
Cell Biology ◽  
Family Members ◽  
Membrane Association ◽  
Rab Gtpases ◽  
Membrane Compartments ◽  
Membrane Bound ◽  
Lrrk2 Kinase ◽  
Familial Parkinson’S Disease ◽  
Distinct Membrane

LRRK2 kinase mutations cause familial Parkinson’s disease and increased phosphorylation of a subset of Rab GTPases. Rab29 recruits LRRK2 to the trans-Golgi and activates it there, yet some of LRRK2’s major Rab substrates are not on the Golgi. We sought to characterize the cell biology of LRRK2 activation. Unlike other Rab family members, we show that Rab29 binds nucleotide weakly, is poorly prenylated, and is not bound to GDI in the cytosol; nevertheless, Rab29 only activates LRRK2 when it is membrane bound and GTP bound. Mitochondrially anchored, GTP-bound Rab29 is both a LRRK2 substrate and activator, and it drives accumulation of active LRRK2 and phosphorylated Rab10 on mitochondria. Importantly, mitochondrially anchored LRRK2 is much less capable of phosphorylating plasma membrane–anchored Rab10 than soluble LRRK2. These data support a model in which LRRK2 associates with and dissociates from distinct membrane compartments to phosphorylate Rab substrates; if anchored, LRRK2 can modify misdelivered Rab substrates that then become trapped there because GDI cannot retrieve them.

scholarly journals RAB8, RAB10 and RILPL1 contribute to both LRRK2 kinase–mediated centrosomal cohesion and ciliogenesis deficits

2019 ◽  
Vol 28 (21) ◽  
pp. 3552-3568 ◽  
Author(s):  
Antonio Jesús Lara Ordónez ◽  
Belén Fernández ◽  
Elena Fdez ◽  
María Romo-Lozano ◽  
Jesús Madero-Pérez ◽  
...  
Keyword(s):  
Rab Gtpases ◽  
Common Cause ◽  
Increase Risk ◽  
Dividing Cells ◽  
Mutant Lrrk2 ◽  
Lrrk2 Kinase ◽  
Relevant Context ◽  
Primary Astrocytes ◽  
Peripheral Cells ◽  
Familial Parkinson’S Disease

Abstract Mutations in the LRRK2 kinase are the most common cause of familial Parkinson’s disease, and variants increase risk for the sporadic form of the disease. LRRK2 phosphorylates multiple RAB GTPases including RAB8A and RAB10. Phosphorylated RAB10 is recruited to centrosome-localized RILPL1, which may interfere with ciliogenesis in a disease-relevant context. Our previous studies indicate that the centrosomal accumulation of phosphorylated RAB8A causes centrosomal cohesion deficits in dividing cells, including in peripheral patient-derived cells. Here, we show that both RAB8 and RAB10 contribute to the centrosomal cohesion deficits. Pathogenic LRRK2 causes the centrosomal accumulation not only of phosho-RAB8 but also of phospho-RAB10, and the effects on centrosomal cohesion are dependent on RAB8, RAB10 and RILPL1. Conversely, the pathogenic LRRK2-mediated ciliogenesis defects correlate with the centrosomal accumulation of both phospho-RAB8 and phospho-RAB10. LRRK2-mediated centrosomal cohesion and ciliogenesis alterations are observed in patient-derived peripheral cells, as well as in primary astrocytes from mutant LRRK2 mice, and are reverted upon LRRK2 kinase inhibition. These data suggest that the LRRK2-mediated centrosomal cohesion and ciliogenesis defects are distinct cellular readouts of the same underlying phospho-RAB8/RAB10/RILPL1 nexus and highlight the possibility that either centrosomal cohesion and/or ciliogenesis alterations may serve as cellular biomarkers for LRRK2-related PD.

A model for Rab GTPase localization

2005 ◽  
Vol 33 (4) ◽  
pp. 627-630 ◽  
Author(s):  
S. Pfeffer
Keyword(s):  
Steady State ◽  
Cellular Localization ◽  
Rab Gtpase ◽  
Rab Proteins ◽  
Rab Gtpases ◽  
Macromolecular Assemblies ◽  
Complex Interactions ◽  
Membrane Compartment ◽  
Membrane Bound ◽  
Distinct Membrane

The human genome encodes almost 70 Rab GTPases. These proteins are C-terminally geranylgeranylated and are localized to the surfaces of distinct membrane-bound compartments in eukaryotic cells. This mini review presents a working model for how Rabs achieve and maintain their steady-state localizations. Data from a number of laboratories suggest that Rabs participate in the generation of macromolecular assemblies that generate functional microdomains within a given membrane compartment. Our data suggest that these complex interactions are important for the cellular localization of Rab proteins at steady state.

Targeting Rab GTPases to distinct membrane compartments

2004 ◽  
Vol 5 (11) ◽  
pp. 886-896 ◽  
Author(s):  
Suzanne Pfeffer ◽  
Dikran Aivazian
Keyword(s):  
Rab Gtpases ◽  
Membrane Compartments ◽  
Distinct Membrane

scholarly journals Novel roles for mammalian septins: from vesicle trafficking to oncogenesis

2001 ◽  
Vol 114 (5) ◽  
pp. 839-844 ◽  
Author(s):  
B. Kartmann ◽  
D. Roth
Keyword(s):  
Plasma Membrane ◽  
Recent Work ◽  
Cell Biology ◽  
Vesicle Trafficking ◽  
Family Members ◽  
Protein Family ◽  
Cellular Functions ◽  
Cellular Processes ◽  
Sequence Homologies ◽  
Genome Analyses

In recent years a convergence of various aspects of cell biology has become apparent, and yet investigators are only beginning to grasp the underlying unifying mechanisms. Among the proteins that participate in diverse aspects of cell biology are the septins. These are a group of novel GTPase proteins that are broadly distributed in many eukaryotes except plants. Although septins were originally identified as a protein family involved in cytokinesis in yeast, recent advances in the field have now ascribed additional functions to these proteins. In particular, the number of known mammalian septin family members has increased dramatically as more data has become available through genome analyses. We suggest a classification for the mammalian septins based on the sequence homologies in their highly divergent N- and C-termini. Recent work suggests novel functions for septins in vesicle trafficking, oncogenesis and compartmentalization of the plasma membrane. Given the ability of the septins to bind GTP and phosphatidylinositol 4,5-bisphosphate in a mutually exclusive manner, these proteins might be crucial elements for the spatial and/or temporal control of diverse cellular functions. As the functions of the septins become unraveled, our understanding of seemingly different cellular processes may move a step further.

Microcompartments within the yeast plasma membrane

2013 ◽  
Vol 394 (2) ◽  
pp. 189-202 ◽  
Author(s):  
Hans Merzendorfer ◽  
Jürgen J. Heinisch
Keyword(s):  
Plasma Membrane ◽  
Cell Biology ◽  
Model Organism ◽  
Dynamic Interaction ◽  
Eukaryotic Cells ◽  
Classical Concept ◽  
Yeast Plasma Membrane ◽  
Bud Neck ◽  
Microscopic Studies ◽  
Membrane Compartments

Abstract Recent research in cell biology makes it increasingly clear that the classical concept of compartmentation of eukaryotic cells into different organelles performing distinct functions has to be extended by microcompartmentation, i.e., the dynamic interaction of proteins, sugars, and lipids at a suborganellar level, which contributes significantly to a proper physiology. As different membrane compartments (MCs) have been described in the yeast plasma membrane, such as those defined by Can1 and Pma1 (MCCs and MCPs), Saccharomyces cerevisiae can serve as a model organism, which is amenable to genetic, biochemical, and microscopic studies. In this review, we compare the specialized microcompartment of the yeast bud neck with other plasma membrane substructures, focusing on eisosomes, cell wall integrity-sensing units, and chitin-synthesizing complexes. Together, they ensure a proper cell division at the end of mitosis, an intricately regulated process, which is essential for the survival and proliferation not only of fungal, but of all eukaryotic cells.

scholarly journals The Cell Biology of LRRK2 in Parkinson’s Disease

2021 ◽  
Author(s):  
Ahsan Usmani ◽  
Farbod Shavarebi ◽  
Annie Hiniker
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Cell Biology ◽  
Kinase Inhibitor ◽  
Genetic Model ◽  
Point Mutations ◽  
Model Systems ◽  
Rab Gtpases ◽  
Recent Developments ◽  
Lrrk2 Kinase

Point mutations in Leucine-rich repeat kinase 2 (LRRK2) are the most common cause of familial Parkinson’s disease (PD) and are implicated in a significant portion of apparently sporadic PD. Clinically, LRRK2-driven PD is indistinguishable from sporadic PD, making it an attractive genetic model for the much more common sporadic PD. In this review, we highlight recent advances in understanding LRRK2's subcellular functions using LRRK2-PD models, while also considering some of the limitations of these model systems. Recent developments of particular importance include new evidence of key LRRK2 functions in the endolysosomal system and LRRK2’s regulation of and by Rab GTPases. Additionally, LRRK2's interaction with the cytoskeleton allowed elucidation of the LRRK2 structure and appears relevant to LRRK2 protein degradation and LRRK2 kinase inhibitor therapies. We further discuss how LRRK2's interactions with other PD-driving genes, such as VPS35, GCase, and α-synuclein, may highlight cellular pathways more broadly disrupted in PD.

scholarly journals TIP47 is a key effector for Rab9 localization

2006 ◽  
Vol 173 (6) ◽  
pp. 917-926 ◽  
Author(s):  
Dikran Aivazian ◽  
Ramon L. Serrano ◽  
Suzanne Pfeffer
Keyword(s):  
Steady State ◽  
Human Genome ◽  
Effector Proteins ◽  
Rab Gtpases ◽  
Interacting Protein ◽  
Cellular Concentration ◽  
Membrane Compartments ◽  
Effector Binding ◽  
Distinct Membrane

The human genome encodes ∼70 Rab GTPases that localize to the surfaces of distinct membrane compartments. To investigate the mechanism of Rab localization, chimeras containing heterologous Rab hypervariable domains were generated, and their ability to bind seven Rab effectors was quantified. Two chimeras could bind effectors for two distinctly localized Rabs; a Rab5/9 hybrid bound both Rab5 and Rab9 effectors, and a Rab1/9 hybrid bound to certain Rab1 and Rab9 effectors. These unusual chimeras permitted a test of the importance of effector binding for Rab localization. In both cases, changing the cellular concentration of a key Rab9 effector, which is called tail-interacting protein of 47 kD, moved a fraction of the proteins from their parental Rab localization to that of Rab9. Thus, relative concentrations of certain competing effectors could determine a chimera's localization. These data confirm the importance of effector interactions for Rab9 localization, and support a model in which effector proteins rely on Rabs as much as Rabs rely on effectors to achieve their correct steady state localizations.

scholarly journals Insulin activates the plasma-membrane and dense-vesicle cyclic AMP phosphodiesterase in hepatocytes by distinct routes

1983 ◽  
Vol 216 (1) ◽  
pp. 245-248 ◽  
Author(s):  
S R Wilson ◽  
A V Wallace ◽  
M D Houslay
Keyword(s):  
Plasma Membrane ◽  
Cyclic Amp ◽  
Insulin Receptor ◽  
Proteinase Inhibitors ◽  
Atp Depletion ◽  
Peripheral Plasma ◽  
Membrane Enzyme ◽  
Membrane Bound ◽  
Subsequent Activation ◽  
Distinct Membrane

Insulin elicits the activation of two distinct membrane-bound cyclic AMP phosphodiesterases when incubated at 37 degrees C for 5 min with intact hepatocytes: the ‘dense-vesicle’ enzyme and the peripheral-plasma-membrane enzyme. In hepatocytes the lysosomotropic agents chloroquine, methylamine and NH4Cl, as well as intracellular ATP depletion elicited by fructose or incubation with insulin at 22 degrees C, blocks selectively the activation of the ‘dense-vesicle’ enzyme. Incubation of hepatocytes with bacitracin, leupeptin and a variety of proteinase inhibitors failed to affect insulin's activation of these two cyclic AMP phosphodiesterases by distinct routes. It is suggested that activation of the ‘dense-vesicle’ enzyme occurs through a pathway triggered by the endocytosis, processing and recycling of the insulin receptor. This might involve the delivery, with subsequent activation, of a latent phosphodiesterase into this fraction.

scholarly journals Insulin and glucagon regulate the activation of two distinct membrane-bound cyclic AMP phosphodiesterases in hepatocytes

1983 ◽  
Vol 214 (1) ◽  
pp. 99-110 ◽  
Author(s):  
C M Heyworth ◽  
A V Wallace ◽  
M D Houslay
Keyword(s):  
Plasma Membrane ◽  
Cyclic Amp ◽  
Cholera Toxin ◽  
Regulatory Protein ◽  
Subcellular Fractionation ◽  
Dibutyryl Cyclic Amp ◽  
Membrane Enzyme ◽  
Membrane Bound ◽  
Guanine Nucleotide Regulatory Protein ◽  
Distinct Membrane

Glucagon (10 nM) caused a transient elevation of intracellular cyclic AMP concentrations, which reached a peak in around 5 min, and slowly returned to basal values in around 30 min. When 1 mM-3-isobutyl-1-methylxanthine (IBMX) was present, this process yielded a Ka of 1 nM for glucagon. The addition of insulin (10 nM) after 5 min exposure to glucagon (10 nM) caused intracellular cyclic AMP concentrations to fall dramatically, attaining basal values within 10 min. The regulation of this process was dose-dependent, exhibiting a Ka of 0.4 nM for insulin. If insulin and glucagon were added together to hepatocytes, then insulin decreased the magnitude of the cyclic AMP response to glucagon. IBMX (1 mM) prevented insulin antagonizing the action of glucagon in both of these instances. A gentle homogenization procedure followed by a rapid subcellular fractionation of hepatocytes on a Percoll gradient was developed. This was used to resolve subcellular membrane fractions and to identify cyclic AMP phosphodiesterase activity in both membrane and cytosol fractions. Glucagon and insulin only affected the activity of two distinct membrane-bound species, a plasma-membrane enzyme and a ‘dense vesicle’ enzyme. Glucagon (10 nM), insulin (10 nM), IBMX (1 mM), dibutyryl cyclic AMP (10 microM) and cholera toxin (1 microgram/ml) all elicited the activation of the ‘dense vesicle’ enzyme. The plasma-membrane enzyme was not activated by glucagon, IBMX or dibutyryl cyclic AMP, although insulin and cholera toxin both led to its activation. The degree of activation of the plasma-membrane enzyme produced by insulin was increased in the presence of IBMX or dibutyryl cyclic AMP. Glucagon pretreatment (5 min) of hepatocytes blocked the ability of insulin to activate the plasma-membrane enzyme. The activity state of these phosphodiesterases is discussed in relation to the observed changes in intracellular cyclic AMP concentrations. It is suggested that insulin exerts its action on the plasma-membrane phosphodiesterase through a mechanism involving a guanine nucleotide-regulatory protein.

Sign in / Sign up

Export Citation Format

Share Document