Inverted Formin 2 Regulates Actin Dynamics by Antagonizing Rho/Diaphanous-related Formin Signaling

*Division of Nephrology, Department of Medicine, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts
Journal of the American Society of Nephrology (Impact Factor: 9.34). 04/2013; 24(6). DOI: 10.1681/ASN.2012080834
Source: PubMed


Mutations in inverted formin 2 INF2 are a common cause of familial FSGS. INF2 interacts with diaphanous-related formins (mDia) and antagonizes mDia-mediated actin polymerization in response to active Rho signaling, suggesting that dysregulation of these pathways may mediate the development of INF2-related FSGS. However, the precise mechanisms by which INF2 regulates actin-dependent podocyte behavior remain largely unknown. Here, we investigated the possible role of INF2 in both lamellipodia-associated actin dynamics and actin-dependent slit diaphragm (SD) protein trafficking by manipulating the expression of INF2 and the activity of Rho/mDia signaling in cultured podocytes. Activation of mDia in the absence of INF2 led to defective formation of lamellipodia and abnormal SD trafficking. Effects of mutations disrupting the INF2-mDia interaction suggested the specificity of the mDia-antagonizing effect of INF2 in maintaining the lamellipodium. Furthermore, we found that SD trafficking requires INF2 interaction with lipid raft components. In summary, INF2 regulates lamellipodial actin dynamics and the trafficking of slit diaphragm proteins by opposing Rho/mDia-mediated actin polymerization. Thus, in podocytes, INF2 appears to be an important modulator of actin-dependent behaviors that are under the control of Rho/mDia signaling.

11 Reads
  • Source
    • "In cultured cells, we have found that this prevents the dampening effect of INF2 on Rho/Dia signaling, leading to imbalanced actin dynamics, perturbation of actin polymerization, and disruption of actin based cellular remodeling processes (e.g. lamellipodia formation and trafficking to the membrane of cultured podocytes (Sun et al., 2013)). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Mutations in Inverted Formin 2 (INF2), a diaphanous formin family protein that regulates actin cytoskeleton dynamics, cause focal segmental glomerulosclerosis (FSGS) and Charcot-Marie-Tooth Disease (CMT) in humans. In addition to directly remodeling actin filaments in vitro, we have shown that INF2 regulates intracellular actin dynamics and actin dependent cellular behavior by opposing Rhoa/Dia signaling. As a step towards a better understanding of the human kidney disease, we wanted to explore the relevance of these findings to the in vivo situation. We used dose dependent knockdown of INF2 to first define an in vivo model and establish an overt glomerular phenotype in zebrafish. This simple assay was validated by rescue with wild type INF2 confirming the specificity of the findings. The edema, podocyte dysfunction, and an altered glomerular filtration barrier observed in the zebrafish pronephros correlate with mistrafficking of glomerular slit diaphragm proteins, defective slit-diaphragm signaling, and disinhibited diaphanous formin (mDia) activity. In contrast to wild-type human INF2, INF2 mutants associated with kidney disease fail to rescue the zINF2 morphant phenotype. Of particular interest, this INF2 knockdown phenotype is also rescued by loss of either RhoA or Dia2. This simple assay allows the demonstration that INF2 functions, at least in part, to modulate Dia-mediated Rho signaling, and that disease causing mutations specifically impair this regulatory function. These data support a model in which disease-associated diaphanous inhibitory domain (DID) mutants in INF2 interfere with its binding to and inhibition of Dia, leading to uncontrolled Rho/Dia signaling and perturbed actin dynamics. Methods to fine tune Rho signaling in the glomerulus may lead to new approaches to therapy in humans.
    12/2014; 1(2-3):107-115. DOI:10.1016/j.ebiom.2014.11.009
  • [Show abstract] [Hide abstract]
    ABSTRACT: Nephrotic syndrome, characterized by massive proteinuria, hypoalbuminemia and edema, is one of the most common kidney diseases in children. Although glucocorticoids (GCs), the mainstay of therapy for over 50 years, are effective in most children, more than 20% develop GC resistant nephrotic syndrome (SRNS), among whom focal segmental glomerular sclerosis (FSGS) is a frequent pathological outcome and the cause of endstage renal disease with a prevalence of 4% in the USA. Despite its clinical importance, the molecular basis of SRNS is unknown. In recent years, researchers have not only gained a new understanding of the roles of structural and functional abnormalities in GC receptors (GRs) in GC resistance, but have also gradually discovered close relationships between GC resistance in idiopathic nephrotic syndrome and podocyte-related molecules, like slit diaphragm (SD) molecules and so on. Here we mainly discussed these molecules and their physiological as well as pathological effects, including nephrin, podocin, CD2-associated protein (CD2AP), a-actinin-4, transient receptor potential cation channel 6 (TRPC6), phospholipase C epsilon-1 (PLCe1), Wilms' tumor suppressor gene 1 (WT1), Lmx1b, LAMB2, myosin 1e (MOY1E) and inverted formin 2 (IFN2). Mitochondrial cytopathies are also involved in GC resistance and well-reviewed [1, 2], which will not be discussed in detail in this review. To those SRNS without any genetic defects, immunological disturbances are always involved and should be stressed. In this article, recent progress in research on the mechanisms of GC resistance in idiopathic nephrotic syndrome is reviewed.
    Kidney and Blood Pressure Research 09/2013; 37(4-5):360-378. DOI:10.1159/000350163 · 2.12 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Members of the Diaphanous (Dia) protein family are key regulators of fundamental actin driven cellular processes, which are conserved from yeast to humans. Researchers have uncovered diverse physiological roles in cell morphology, cell motility, cell polarity, and cell division, which are involved in shaping cells into tissues and organs. The identification of numerous binding partners led to substantial progress in our understanding of the differential functions of Dia proteins. Genetic approaches and new microscopy techniques allow important new insights into their localization, activity, and molecular principles of regulation.
    Communicative & integrative biology 11/2013; 6(6):e27634. DOI:10.4161/cib.27634
Show more