S-Nitrosylation of RhoGAP Myosin9A Is Altered in Advanced Diabetic Kidney Disease

Overview

Journal Front Med (Lausanne)

Specialty General Medicine

Date 2021 Aug 2

PMID 34336885

Citations 3

Authors

Qi Li

Delma Veron

Alda Tufro

Affiliations

Soon will be listed here.

Abstract

The molecular pathogenesis of diabetic kidney disease progression is complex and remains unresolved. Rho-GAP was recently identified as a novel podocyte protein and a candidate gene for monogenic FSGS. Myo9A involvement in diabetic kidney disease has been suggested. Here, we examined the effect of diabetic milieu on Myo9A expression and . We determined that Myo9A undergoes S-nitrosylation, a post-translational modification dependent on nitric oxide (NO) availability. Diabetic mice with nodular glomerulosclerosis and severe proteinuria associated with doxycycline-induced, podocyte-specific gain-of-function showed markedly decreased glomerular Myo9A expression and S-nitrosylation, as compared to uninduced diabetic mice. Immortalized mouse podocytes exposed to high glucose revealed decreased expression, assessed by qPCR, immunoblot and immunocytochemistry, and reduced Myo9A S-nitrosylation (SNO-Myo9A), assessed by proximity link assay and biotin switch test, functionally resulting in abnormal podocyte migration. These defects were abrogated by exposure to a NO donor and were not due to hyperosmolarity. Our data demonstrate that high-glucose induced decrease of both expression and SNO-Myo9A is regulated by NO availability. We detected S-nitrosylation of Myo9A interacting proteins RhoA and actin, which was also altered by high glucose and NO dependent. RhoA activity inversely related to SNO-RhoA. Collectively, data suggest that dysregulation of SNO-Myo9A, SNO-RhoA and SNO-actin may contribute to the pathogenesis of advanced diabetic kidney disease and may be amenable to therapeutic targeting.

Citing Articles

Redox regulation in diabetic kidney disease.

Daehn I, Ekperikpe U, Stadler K Am J Physiol Renal Physiol. 2023; 325(2):F135-F149.

PMID: 37262088 PMC: 10393330. DOI: 10.1152/ajprenal.00047.2023.

Nitric-Oxide-Mediated Signaling in Podocyte Pathophysiology.

Semenikhina M, Stefanenko M, Spires D, Ilatovskaya D, Palygin O Biomolecules. 2022; 12(6).

PMID: 35740870 PMC: 9221338. DOI: 10.3390/biom12060745.

Podocyte Knockdown Induces Diffuse Glomerulosclerosis in Diabetic and in Knockout Mice.

Veron D, Aggarwal P, Li Q, Moeckel G, Kashgarian M, Tufro A Front Pharmacol. 2022; 12:788886.

PMID: 35280251 PMC: 8906751. DOI: 10.3389/fphar.2021.788886.

References

Tufro A, Veron D . VEGF and podocytes in diabetic nephropathy. Semin Nephrol. 2012; 32(4):385-93. PMC: 3438453. DOI: 10.1016/j.semnephrol.2012.06.010. View

Zhang Y, Peng F, Gao B, Ingram A, Krepinsky J . High glucose-induced RhoA activation requires caveolae and PKCβ1-mediated ROS generation. Am J Physiol Renal Physiol. 2011; 302(1):F159-72. DOI: 10.1152/ajprenal.00749.2010. View

Iwakiri Y, Satoh A, Chatterjee S, Toomre D, Chalouni C, Fulton D . Nitric oxide synthase generates nitric oxide locally to regulate compartmentalized protein S-nitrosylation and protein trafficking. Proc Natl Acad Sci U S A. 2006; 103(52):19777-82. PMC: 1750883. DOI: 10.1073/pnas.0605907103. View

Evangelista A, Kohr M, Murphy E . S-nitrosylation: specificity, occupancy, and interaction with other post-translational modifications. Antioxid Redox Signal. 2012; 19(11):1209-19. PMC: 3785808. DOI: 10.1089/ars.2012.5056. View

Horenberg A, Houghton A, Pandey S, Seshadri V, Guilford W . S-nitrosylation of cytoskeletal proteins. Cytoskeleton (Hoboken). 2019; 76(3):243-253. DOI: 10.1002/cm.21520. View

Stomberski C, Hess D, Stamler J . Protein S-Nitrosylation: Determinants of Specificity and Enzymatic Regulation of S-Nitrosothiol-Based Signaling. Antioxid Redox Signal. 2017; 30(10):1331-1351. PMC: 6391618. DOI: 10.1089/ars.2017.7403. View

Tian D, Jacobo S, Billing D, Rozkalne A, Gage S, Anagnostou T . Antagonistic regulation of actin dynamics and cell motility by TRPC5 and TRPC6 channels. Sci Signal. 2010; 3(145):ra77. PMC: 3071756. DOI: 10.1126/scisignal.2001200. View

Gorman S, Haider N, Grieshammer U, Swiderski R, Kim E, Welch J . The cloning and developmental expression of unconventional myosin IXA (MYO9A) a gene in the Bardet-Biedl syndrome (BBS4) region at chromosome 15q22-q23. Genomics. 1999; 59(2):150-60. DOI: 10.1006/geno.1999.5867. View

Heo J, Raines K, Mocanu V, Campbell S . Redox regulation of RhoA. Biochemistry. 2006; 45(48):14481-9. DOI: 10.1021/bi0610101. View

10.

Ingelfinger J, Rosen C . Clinical Credence - SGLT2 Inhibitors, Diabetes, and Chronic Kidney Disease. N Engl J Med. 2019; 380(24):2371-2373. DOI: 10.1056/NEJMe1904740. View

11.

Soderberg O, Gullberg M, Jarvius M, Ridderstrale K, Leuchowius K, Jarvius J . Direct observation of individual endogenous protein complexes in situ by proximity ligation. Nat Methods. 2006; 3(12):995-1000. DOI: 10.1038/nmeth947. View

12.

Hanley P, Vollmer V, Bahler M . Class IX Myosins: Motorized RhoGAP Signaling Molecules. Adv Exp Med Biol. 2020; 1239:381-389. DOI: 10.1007/978-3-030-38062-5_16. View

13.

Veron D, Reidy K, Bertuccio C, Teichman J, Villegas G, Jimenez J . Overexpression of VEGF-A in podocytes of adult mice causes glomerular disease. Kidney Int. 2010; 77(11):989-99. DOI: 10.1038/ki.2010.64. View

14.

Erwin P, Lin A, Golan D, Michel T . Receptor-regulated dynamic S-nitrosylation of endothelial nitric-oxide synthase in vascular endothelial cells. J Biol Chem. 2005; 280(20):19888-94. DOI: 10.1074/jbc.M413058200. View

15.

Yi F, Kong R, Ren J, Zhu L, Lou J, Wu J . Noncanonical Myo9b-RhoGAP Accelerates RhoA GTP Hydrolysis by a Dual-Arginine-Finger Mechanism. J Mol Biol. 2016; 428(15):3043-57. PMC: 5714546. DOI: 10.1016/j.jmb.2016.06.014. View

16.

Sonneveld R, van der Vlag J, Baltissen M, Verkaart S, Wetzels J, Berden J . Glucose specifically regulates TRPC6 expression in the podocyte in an AngII-dependent manner. Am J Pathol. 2014; 184(6):1715-26. DOI: 10.1016/j.ajpath.2014.02.008. View

17.

Warren A, Knudsen S, Cooper M . Diabetic nephropathy: an insight into molecular mechanisms and emerging therapies. Expert Opin Ther Targets. 2019; 23(7):579-591. DOI: 10.1080/14728222.2019.1624721. View

18.

Thom S, Bhopale V, Mancini D, Milovanova T . Actin S-nitrosylation inhibits neutrophil beta2 integrin function. J Biol Chem. 2008; 283(16):10822-34. DOI: 10.1074/jbc.M709200200. View

19.

Peng H, Luo P, Li Y, Wang C, Liu X, Ye Z . Simvastatin alleviates hyperpermeability of glomerular endothelial cells in early-stage diabetic nephropathy by inhibition of RhoA/ROCK1. PLoS One. 2013; 8(11):e80009. PMC: 3828237. DOI: 10.1371/journal.pone.0080009. View

20.

Veron D, Bertuccio C, Marlier A, Reidy K, Garcia A, Jimenez J . Podocyte vascular endothelial growth factor (Vegf₁₆₄) overexpression causes severe nodular glomerulosclerosis in a mouse model of type 1 diabetes. Diabetologia. 2011; 54(5):1227-41. PMC: 3397150. DOI: 10.1007/s00125-010-2034-z. View