Encyclopedia of Signaling Molecules

2018 Edition
| Editors: Sangdun Choi

RGS10

Reference work entry
DOI: https://doi.org/10.1007/978-3-319-67199-4_101851

Historical Background

The regulator of G protein signaling 10 (RGS10) protein is a regulatory molecule that belongs to a larger family of RGS proteins responsible for altering cellular signaling. Generally, RGS proteins are negative regulators of signal transduction pathways mediated by heterotrimeric G proteins. This function allows RGS proteins to control the duration and amplification of signaling activity, ultimately operating as shut-off switches. RGS proteins deactivate G protein subunits by serving as GTPase activating proteins (GAPs), which enhance the intrinsic GTPase activity of the active, GTP-bound Gα subunit and return G proteins to their inactive, GDP-bound form. The result is an acceleration of the deactivation of G protein signaling.

Protein Structure and Function

All RGS proteins share a conserved 120 amino acid helical RGS domain that is responsible for GAP activity. Based on sequence similarity of the RGS domain, RGS10 is grouped in the RGS 12 subfamily which also includes the proteins RGS12 and RGS14 (Alexander et al. 2015). Despite the similarity of their RGS domains, RGS10 (20 kDa) is much smaller than both RGS14 (61 kDa) and RGS12 (156 kDa), as RGS10 lacks the regulatory and multifunctional domains found in RGS12 and RGS14. Indeed, RGS10 is one of the smallest members of the entire RGS protein family. Aside from its RGS domain, RGS10 contains only short N-terminal and C-terminal extensions that lack any defined domain structure but may be involved in subcellular targeting (see below).

Gα subunits of heterotrimeric G proteins possess intrinsic GTPase activity toward the γ-phosphate of GTP and undergo significant conformational changes during the transition from the active GTP-bound form to the inactive GDP-bound form. RGS proteins mediate their GAP activity by selectively binding and stabilizing the transition state nucleotide-bound form of Gα subunit, which facilitates and accelerates hydrolysis (Fig. 1a). This mechanism is distinct from that of the GAPs for small GTPases of the Ras superfamily, which directly contribute to GTP hydrolysis by inserting either an arginine or asparagine in the GTPase active site (Ahmadian et al. 1997; Li et al. 2004). RGS domains interact with flexible switch regions of Gα protein subunits to stabilize the transition state but do not make contacts with the active site. RGS10, and all R12 RGS subfamily members, selectively bind and deactivate Gi family Gα subunits, including Gαi, Gαo, and Gαz, with very low affinity for Gαq (Hunt et al. 1996; Watson et al. 1996; Popov et al. 1997; Soundararajan et al. 2008; Taylor et al. 2016).
RGS10, Fig. 1

Canonical function and localization of the RGS10 protein. (a) After G protein-coupled receptor signaling activation, RGS10 serves as a GAP by potently stimulating the hydrolysis of GTP bound to GαZ or Gαi3, terminating furthur signaling. This deactivation of G protein subunits is the canonical function of RGS proteins. (b) Interestingly, RGS10 is localized to the nucleus. This action occurs through the phosphorylation of serine 168 by PKA, thereby causing the translocation of RGS10

The structure of RGS10 and its interaction with Gαi subunits has been defined in detail (Soundararajan et al. 2008; Taylor et al. 2016). RGS10 contains the conserved RGS domain structure consisting of nine α helices, although Soundararajan and colleagues demonstrated that RGS10 possesses a shorter αVI helix and an extended αV–αVI loop compared to RGS domains of R4, R7, and RZ subfamilies. The RGS10-Gαi3 complex is generally consistent with G-protein complexes with other RGS subfamily members, but differences were again observed in the αVI helix. For example, a conserved RGS domain arginine residue in helix αVI that makes contacts with the Gα switch III region in complexes formed with members of the R4 family did not make contacts with Gαi3 in the RGS10 complex (Soundararajan et al. 2008). Finally, the GAP activity of RGS10 is under complex regulation by palmitoylation of a conserved cysteine in the RGS domain. Palmitoylation of this residue inhibits RGS10 single turnover GTPase activity but substantially potentiates steady-state GAP activity in proteoliposome reconstituted receptor-G protein complexes (Tu et al. 1997, 1999).

Localization

In spite of the plasma membrane localization of their canonical G protein targets, many RGS proteins are localized to the nucleus. This localization may serve as a cellular storage compartment to sequester RGS proteins and regulate their GAP activity, or may allow RGS proteins to regulate undefined nuclear targets (Hollinger and Hepler 2002). Several studies have demonstrated RGS10 localization to the nucleus and cytoplasm, with minimal plasma membrane localization (Chatterjee and Fisher 2000; Lee et al. 2008). Putative nuclear localization sequences have been identified in the RGS10 N- and C-termini but have not been functionally validated. Further, the nuclear localization of RGS10 is dynamically regulated by cellular activation (Lee et al. 2008) and through cyclic AMP-dependent protein kinase A mediated phosphorylation of RGS10 on serine 168 at its C terminus (Burgon et al. 2001) (Fig. 1b). Further implications and interpretations of this noncanonical localization by a G protein regulatory molecule suggest unknown and/or unappreciated roles for RGS10 outside of canonical understanding.

Gene Organization

RGS10 is conserved in tetrapods including mammals, birds, and amphibians and is encoded by a single gene (Rgs10) on chromosome 10 in Homo sapiens (10q26.11). The mouse Rgs10 gene contains two transcriptional start sites, yielding two transcript isoforms with distinct first exons, and four common exons (Haller et al. 2002; Lee and Tansey 2015). Mouse transcripts mRgs10-1 and mRgs10-2 correspond to human transcripts with conserved exon structure and encoding highly conserved human RGS10 protein isoforms RGS10a (181 aa) and RGS10b (167 aa). A third human transcript (173 aa) has been proposed, resulting from an alternate transcriptional start site upstream of the first shared exon (Hunt et al. 1996), but the relevance of this transcript in mouse cells is unclear (Haller et al. 2002). Rgs10-1/RGS10a is the predominant form expressed in ovarian cancer cells, osteoclasts, microglia, and other immune cells (Yang and Li 2007; Yang et al. 2007; Ali et al. 2013; Lee and Tansey 2015).

Epigenetic Regulation of Expression

Cells control the expression of the Rgs10 gene epigenetically. In chemoresistant ovarian cancer cells, suppression of the RGS10 protein occurs due to both histone deacetylation and DNA hypermethylation, which are common chromatin modifications that synergistically regulate many genes. For example, knocking down the expression of either histone deacetylase 1 or DNA methyltransferase 1 significantly increases the expression of Rgs10 (Cacan et al. 2014). In addition, pharmacological inhibition of DNA methyltransferase activity increases the expression of Rgs10 transcription in neural progenitor cells (Tuggle et al. 2014). In contrast, in activated microglia Rgs10 is suppressed by histone deacetylation while DNA methylation does not have a significant effect. Thus, distinct cell types use distinct epigenetic mechanisms to suppress RGS10 levels, which will indirectly amplify the G protein signaling pathways that are targeted by RGS10.

Given that Rgs10 expression is dynamically regulated, Rgs10 expression levels or epigenetic marks may be useful biomarkers. A genome-wide study examining freely circulating DNA in the blood of patients with hepatocellular carcinoma using a technique called methylated CpG tandem amplification and sequencing (MCTA-Seq) found hypermethylated CpG islands of RGS10 in circulation. The study used an unbiased method to detect aberrant DNA methylation changes and showed that only 2.3% of hypermethylated CpG islands were sufficient biomarkers for detecting hepatocellular carcinoma in blood. Most importantly, RGS10 was among only four biomarkers that fit tumor-specific criteria used in this study (Wen et al. 2015) Thus, epigenetically modified RGS10 may have the potential to serve as a specific biomarker for the early stage detection of hepatocellular carcinoma.

Signaling in Ovarian Cancer Chemoresistance

Comparison of transcript expression datasets between ovarian cancer cells that were either chemosensitive or chemoresistant demonstrated that Rgs10 expression is suppressed with the development of drug resistance. The significance of this change in expression was demonstrated by the observation that AKT signaling was increased following suppression of RGS10 (Hooks et al. 2010). Further elucidation of the molecular mechanism demonstrated that activation of the mTOR signaling pathway is increased with RGS10 suppression, as demonstrated by the phosphorylation of 4E-BP1, p70 S6 kinase, eIF2a, AKT, and S6 ribosomal protein. In a noncanonical role, RGS10 was shown to bind to the small GTPase protein Rheb, which binds and activates mTORC1. siRNA inhibition of RGS10 increased the levels of active GTP-bound Rheb, suggesting that RGS10 may negatively regulate the mTOR pathway through Rheb in these cells (Altman et al. 2015) (Fig. 2).
RGS10, Fig. 2

A noncanonical function of RGS10. Biochemical data demonstrated that GRS10 binds to the small GTPase Rheb. This protein shuttles between its GDP- and GTP-bound forms and serves as an mTORC1 activator in its GTP-bound form. Loss of RGS10 leads to an increase in phosphorylation of mTOR (Ser2448) and several of its downstream effector targets: 4E-BP1 (Thr37/46), elF2a (Ser51), S6 ribosomal protein (Ser235/236), and p70S6 kinase (Thr389)

Signaling in Platelets

Platelets respond to multiple GPCR agonists to regulate hemostasis and thrombogenesis. In platelets, RGS10 binds to a complex with the spinophilin scaffolding protein and the SHP-1 protein tyrosine phosphatase to negatively regulate platelet activation until further SHP-1 phosphorylation occurs, which causes the decay of this complex (Ma et al. 2012). In platelets derived from RGS10−/− mice, agonist-induced signaling was hyperresponsive and manifested by platelet aggregation and enhanced hemostasis. In these mice, there was also an observation of increased susceptibility to ischemia due to thrombosis (Hensch et al. 2016). Strikingly, loss of RGS10 expression amplified platelet aggregation in response to activation of receptors coupled to Gi, Gq, and G12 receptors, despite the clear Gi selectivity of RGS10 GAP activity. These studies suggest that RGS10 acts as an integral “braking system” that blunts platelet activation and thrombogenesis, but the mechanism remains undefined.

Signaling in Immune Cells

RGS10 is highly expressed in the brain and immune system, with particularly high expression in microglia, the resident macrophages of the central nervous system. Several studies have demonstrated that microglial RGS10 plays important anti-inflammatory and neuroprotective roles by inhibiting lipopolysaccharide-stimulated production of inflammatory cytokines including TNFα (Lee et al. 2008, 2011). Additional neuroprotective effects are mediated by neuronal RGS10, which increases cell survival and resistance to TNF-induced cytotoxicity (Lee et al. 2012). Together, these results suggest that RGS10 may be protective in neurodegenerative diseases with underlying neuroinflammation, such as Parkinson’s disease. In contrast, RGS10 is implicated in promoting progression of the autoimmune neurodegenerative disease multiple sclerosis (MS), and RGS10−/− display dramatically milder symptoms in a mouse model of MS (Lee et al. 2016). The effect of RGS10 in this model is attributed to its effects on peripheral T cells. RGS10 is also a critical regulator of peripheral macrophages, promoting the anti-inflammatory M2 alternative activation while suppressing the inflammatory M1 activation phenotype (Lee et al. 2013). Thus, RGS10 plays diverse roles in various cells of the immune system and may contribute to multiple neuroinflammatory and immune disorders. In particular, RGS10 may contribute to inflammatory disorders related to aging, as RGS10 expression changes significantly in various immune cells during aging (Kannarkat et al. 2015).

Signaling in Heart Failure

RGS10 protein levels are significantly lower in failing human hearts and in a mouse model of cardiac hypertrophy compared to normal controls, while overexpression of RGS10 suppresses cardiac hypertrophy in cell and animal models. Further, RGS10 inhibits signaling downstream of the Gαq coupled angiotensin II receptor, which induces cardiomyocyte hypertrophy. Thus, RGS10 negatively regulates signaling in this model and inhibits cardiac hypertrophy (Miao et al. 2016).

Signaling in Osteoclast Differentiation

RGS10 is dynamically expressed in bone resorbing osteoclast cells, and RGS10 plays an important role in regulating osteoclast function and differentiation. The RANKL-stimulated signaling pathway for differentiation of osteoclasts induces the production of RGS10. When RGS10 is suppressed in this model, intracellular Ca2+ oscillations are inhibited as well as expression of NFAT2 – the master switch for regulating terminal differentiation of osteoclasts. RGS10−/− mice display impaired osteoclast differentiation and osteopetrosis. This suggests that RGS10 is a key regulator that cooperates with other factors activated by RANKL and is an essential component of this signaling pathway in bone physiology (Yang and Li 2007; Yang et al. 2007).

Summary

RGS10 is an important regulatory protein that negatively regulates signaling events in a diverse range of cell types (Fig. 3). Since the mid 1990s, research has established canonical roles for RGS10 in the regulation of G proteins. Exciting discoveries have been elucidated in recent years; however, the full repertoire of RGS10’s importance as a signaling molecule and biomarker has not yet been completely defined. Suppression of RGS10 is associated with multiple pathological states, and this suppression is a mediating factor of pathology through the hyperactivation of cellular signaling. Further efforts are needed to uncover additional regulatory factors, a more thorough role for RGS10 in pharmacology as well as disease mechanisms. A better basic understanding of its role in these areas would help in the development of future therapies in a variety of ailments.
RGS10, Fig. 3

Biological roles of RGS10. The GAP is involved in many necessary functions, including drug sensitivity, platelet function, neuroprotection of microglia, inhibiting hypertrophy in cardiomycetes and as a key regulator of osteoclasts

References

  1. Ahmadian MR, Stege P, Scheffzek K, Wittinghofer A. Confirmation of the arginine-finger hypothesis for the GAP-stimulated GTP-hydrolysis reaction of Ras. Nat Struct Biol. 1997;4:686–9.PubMedCrossRefGoogle Scholar
  2. Alexander SPH, Kelly E, Marrion N, Peters JA, Benson HE, Faccenda E, et al. The concise guide to pharmacology 2015/16: overview. Br J Pharmacol. 2015;172:5729–43.PubMedPubMedCentralCrossRefGoogle Scholar
  3. Ali MW, Cacan E, Liu Y, Pierce JY, Creasman WT, Murph MM, et al. Transcriptional suppression, DNA methylation, and histone deacetylation of the regulator of G-protein signaling 10 (RGS10) gene in ovarian cancer cells. PLoS One. 2013;8:e60185. doi: 10.1371/journal.pone.0060185.PubMedPubMedCentralCrossRefGoogle Scholar
  4. Altman MK, Alshamrani AA, Jia W, Nguyen HT, Fambrough JM, Tran SK, et al. Suppression of the GTPase-activating protein RGS10 increases Rheb-GTP and mTOR signaling in ovarian cancer cells. Cancer Lett. 2015;369:175–83. doi: 10.1016/j.canlet.2015.08.012.PubMedPubMedCentralCrossRefGoogle Scholar
  5. Burgon PG, Lee WL, Nixon AB, Peralta EG, Casey PJ. Phosphorylation and nuclear translocation of a regulator of G protein signaling (RGS10). J Biol Chem. 2001;276:32828–34. doi: 10.1074/jbc.M100960200.PubMedCrossRefGoogle Scholar
  6. Cacan E, Ali MW, Boyd NH, Hooks SB, Greer SF. Inhibition of HDAC1 and DNMT1 modulate RGS10 expression and decrease ovarian cancer chemoresistance. PLoS One. 2014;9:e87455. doi: 10.1371/journal.pone.0087455.PubMedPubMedCentralCrossRefGoogle Scholar
  7. Chatterjee TK, Fisher RA. Cytoplasmic, nuclear, and golgi localization of RGS proteins. Evidence for N-terminal and RGS domain sequences as intracellular targeting motifs. J Biol Chem. 2000;275:24013–21. doi: 10.1074/jbc.M002082200.PubMedCrossRefGoogle Scholar
  8. Haller C, Fillatreau S, Hoffmann R, Agenes F. Structure, chromosomal localization and expression of the mouse regulator of G-protein signaling10 gene (mRGS10). Gene. 2002;297:39–49.PubMedCrossRefGoogle Scholar
  9. Hensch NR, Karim ZA, Druey KM, Tansey MG, Khasawneh FT. RGS10 negatively regulates platelet activation and thrombogenesis. PLoS One. 2016;11:e0165984. doi: 10.1371/journal.pone.0165984.PubMedPubMedCentralCrossRefGoogle Scholar
  10. Hollinger S, Hepler JR. Cellular regulation of RGS proteins: modulators and integrators of G protein signaling. Pharmacol Rev. 2002;54:527–59.PubMedCrossRefGoogle Scholar
  11. Hooks SB, Callihan P, Altman MK, Hurst JH, Ali MW, Murph MM. Regulators of G-protein signaling RGS10 and RGS17 regulate chemoresistance in ovarian cancer cells. Mol Cancer. 2010;9:289. doi: 10.1186/1476-4598-9-289.PubMedPubMedCentralCrossRefGoogle Scholar
  12. Hunt TW, Fields TA, Casey PJ, Peralta EG. RGS10 is a selective activator of G alpha i GTPase activity. Nature. 1996;383:175–7. doi: 10.1038/383175a0.PubMedCrossRefGoogle Scholar
  13. Kannarkat GT, Lee JK, Ramsey CP, Chung J, Chang J, Porter I, et al. Age-related changes in regulator of G-protein signaling (RGS)-10 expression in peripheral and central immune cells may influence the risk for age-related degeneration. Neurobiol Aging. 2015;36:1982–93. doi: 10.1016/j.neurobiolaging.2015.02.006.PubMedPubMedCentralCrossRefGoogle Scholar
  14. Lee JK, Tansey MG. Physiology of RGS10 in neurons and immune cells. Prog Mol Biol Transl Sci. 2015;133:153–67. doi: 10.1016/bs.pmbts.2015.01.005.PubMedCrossRefGoogle Scholar
  15. Lee JK, McCoy MK, Harms AS, Ruhn KA, Gold SJ, Tansey MG. Regulator of G-protein signaling 10 promotes dopaminergic neuron survival via regulation of the microglial inflammatory response. J Neurosci Off J Soc Neurosci. 2008;28:8517–28. doi: 10.1523/JNEUROSCI.1806-08.2008.CrossRefGoogle Scholar
  16. Lee JK, Chung J, McAlpine FE, Tansey MG. Regulator of G-protein signaling-10 negatively regulates NF-kappaB in microglia and neuroprotects dopaminergic neurons in hemiparkinsonian rats. J Neurosci Off J Soc Neurosci. 2011;31:11879–88. doi: 10.1523/JNEUROSCI.1002-11.2011.CrossRefGoogle Scholar
  17. Lee JK, Chung J, Druey KM, Tansey MG. RGS10 exerts a neuroprotective role through the PKA/c-AMP response-element (CREB) pathway in dopaminergic neuron-like cells. J Neurochem. 2012;122:333–43. doi: 10.1111/j.1471-4159.2012.07780.x.PubMedPubMedCentralCrossRefGoogle Scholar
  18. Lee JK, Chung J, Kannarkat GT, Tansey MG. Critical role of regulator G-protein signaling 10 (RGS10) in modulating macrophage M1/M2 activation. PLoS One. 2013;8:e81785. doi: 10.1371/journal.pone.0081785.PubMedPubMedCentralCrossRefGoogle Scholar
  19. Lee JK, Kannarkat GT, Chung J, Joon Lee H, Graham KL, Tansey MG. RGS10 deficiency ameliorates the severity of disease in experimental autoimmune encephalomyelitis. J Neuroinflammation. 2016;13:24. doi: 10.1186/s12974-016-0491-0.PubMedPubMedCentralCrossRefGoogle Scholar
  20. Li Y, Inoki K, Guan KL. Biochemical and functional characterizations of small GTPase Rheb and TSC2 GAP activity. Mol Cell Biol. 2004;24:7965–75. doi: 10.1128/MCB.24.18.7965-7975.2004.PubMedPubMedCentralCrossRefGoogle Scholar
  21. Ma P, Cierniewska A, Signarvic R, Cieslak M, Kong H, Sinnamon AJ, et al. A newly identified complex of spinophilin and the tyrosine phosphatase, SHP-1, modulates platelet activation by regulating G protein-dependent signaling. Blood. 2012;119:1935–45. doi: 10.1182/blood-2011-10-387910.PubMedPubMedCentralCrossRefGoogle Scholar
  22. Miao R, Lu Y, Xing X, Li Y, Huang Z, Zhong H, et al. Regulator of G-protein signaling 10 negatively regulates cardiac remodeling by blocking mitogen-activated protein kinase-extracellular signal-regulated protein kinase 1/2 signaling. Hypertension. 2016;67:86–98. doi: 10.1161/HYPERTENSIONAHA.115.05957.PubMedCrossRefGoogle Scholar
  23. Popov S, Yu K, Kozasa T, Wilkie TM. The regulators of G protein signaling (RGS) domains of RGS4, RGS10, and GAIP retain GTPase activating protein activity in vitro. Proc Natl Acad Sci USA. 1997;94:7216–20.PubMedPubMedCentralCrossRefGoogle Scholar
  24. Soundararajan M, Willard FS, Kimple AJ, Turnbull AP, Ball LJ, Schoch GA, et al. Structural diversity in the RGS domain and its interaction with heterotrimeric G protein alpha-subunits. Proc Natl Acad Sci USA. 2008;105:6457–62. doi: 10.1073/pnas.0801508105.PubMedPubMedCentralCrossRefGoogle Scholar
  25. Taylor VG, Bommarito PA, Tesmer JJ. Structure of the regulator of G protein signaling 8 (RGS8)-Galphaq complex: molecular basis for Galpha selectivity. J Biol Chem. 2016;291:5138–45. doi: 10.1074/jbc.M115.712075.PubMedPubMedCentralCrossRefGoogle Scholar
  26. Tu Y, Wang J, Ross EM. Inhibition of brain Gz GAP and other RGS proteins by palmitoylation of G protein alpha subunits. Science. 1997;278:1132–5.PubMedCrossRefGoogle Scholar
  27. Tu Y, Popov S, Slaughter C, Ross EM. Palmitoylation of a conserved cysteine in the regulator of G protein signaling (RGS) domain modulates the GTPase-activating activity of RGS4 and RGS10. J Biol Chem. 1999;274:38260–7.PubMedCrossRefGoogle Scholar
  28. Tuggle K, Ali MW, Salazar H, Hooks SB. Regulator of G protein signaling transcript expression in human neural progenitor differentiation: R7 subfamily regulation by DNA methylation. Neurosignals. 2014;22:43–51. doi: 10.1159/000362128.PubMedCrossRefGoogle Scholar
  29. Watson N, Linder ME, Druey KM, Kehrl JH, Blumer KJ. RGS family members: GTPase-activating proteins for heterotrimeric G-protein alpha-subunits. Nature. 1996;383:172–5. doi: 10.1038/383172a0.PubMedCrossRefGoogle Scholar
  30. Wen L, Li J, Guo H, Liu X, Zheng S, Zhang D, et al. Genome-scale detection of hypermethylated CpG islands in circulating cell-free DNA of hepatocellular carcinoma patients. Cell Res. 2015;25:1250–64. doi: 10.1038/cr.2015.126.PubMedPubMedCentralCrossRefGoogle Scholar
  31. Yang S, Li YP. RGS10-null mutation impairs osteoclast differentiation resulting from the loss of [Ca2+]i oscillation regulation. Genes Dev. 2007;21:1803–16. doi: 10.1101/gad.1544107.PubMedPubMedCentralCrossRefGoogle Scholar
  32. Yang S, Chen W, Stashenko P, Li YP. Specificity of RGS10A as a key component in the RANKL signaling mechanism for osteoclast differentiation. J Cell Sci. 2007;120:3362–71. doi: 10.1242/jcs.008300.PubMedPubMedCentralCrossRefGoogle Scholar

Copyright information

© Springer International Publishing AG 2018

Authors and Affiliations

  1. 1.Department of Pharmaceutical and Biomedical SciencesThe University of Georgia, College of PharmacyAthensUSA