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Vanderbilt University
1.
Lu, Lucy Xiangxi.
Phosphoregulation of the Cdc25 phosphatase and its effects on Schizosaccharomyces pombe mitotic entrance and exit.
Degree: PhD, Cell and Developmental Biology, 2012, Vanderbilt University
URL: http://hdl.handle.net/1803/13924 
► Cdk1 kinase dephosphorylation and activation by Cdc25 phosphatase is essential for mitotic entry. Activated Cdk1 phosphorylates Cdc25 and other substrates, further activating Cdc25 to form…
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▼ Cdk1 kinase dephosphorylation and activation by Cdc25 phosphatase is essential for mitotic entry. Activated Cdk1 phosphorylates Cdc25 and other substrates, further activating Cdc25 to form a positive feedback loop that drives the abrupt G2/mitosis switch. Conversely, mitotic exit requires Cdk1 inactivation and reversal of Cdk1 substrate phosphorylation. This is mediated in part by Clp1/Cdc14, a Cdk1 antagonizing phosphatase, which reverses Cdk1 phosphorylation of itself, Cdc25, and other Cdk1 substrates. Thus, Cdc25 phosphoregulation is essential for proper G2-M transition and its contributions to cell cycle control have been modeled based on studies using Xenopus and human cell extracts. Since cell extract systems only approximate in vivo conditions where proteins interact within dynamic cellular environments, here we use Schizosaccharomyces pombe to characterize experimentally and mathematically, the in vivo contributions of Cdk1-mediated phosphorylation of Cdc25 to the mitotic transition. Through comprehensive mapping of Cdk1 phosphosites on Cdc25 and characterization of phosphomutants, we show that Cdc25 hyperphosphorylation by Cdk1 governs Cdc25 catalytic activation, the precision of mitotic entry, and unvarying cell length, but not Cdc25 localization or abundance. We propose a mathematical model that explains Cdc25 regulation by Cdk1 through a distributive and disordered phosphorylation mechanism that ultrasensitively activates Cdc25. We also show that Clp1/Cdc14 dephosphorylation of Cdk1 sites on Cdc25 controls the proper timing of cell division, a mechanism that is likely due to the double negative feedback loop between Clp1/Cdc14 and Cdc25 that controls the abruptness of the mitotic exit switch.
Advisors/Committee Members: Brian E. Wadzinski (committee member), David Cortez (committee member), Laura A. Lee (committee member), William Tansey (Committee Chair).
Subjects/Keywords: Cdk1; phosphatases; kinases; mitosis; bistability; ultrasensitivity; schizosaccharomyces pombe; Cdc14; Clp1; Cdc25; Cdc2
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APA (6th Edition):
Lu, L. X. (2012). Phosphoregulation of the Cdc25 phosphatase and its effects on Schizosaccharomyces pombe mitotic entrance and exit. (Doctoral Dissertation). Vanderbilt University. Retrieved from http://hdl.handle.net/1803/13924
Chicago Manual of Style (16th Edition):
Lu, Lucy Xiangxi. “Phosphoregulation of the Cdc25 phosphatase and its effects on Schizosaccharomyces pombe mitotic entrance and exit.” 2012. Doctoral Dissertation, Vanderbilt University. Accessed January 19, 2021.
http://hdl.handle.net/1803/13924.
MLA Handbook (7th Edition):
Lu, Lucy Xiangxi. “Phosphoregulation of the Cdc25 phosphatase and its effects on Schizosaccharomyces pombe mitotic entrance and exit.” 2012. Web. 19 Jan 2021.
Vancouver:
Lu LX. Phosphoregulation of the Cdc25 phosphatase and its effects on Schizosaccharomyces pombe mitotic entrance and exit. [Internet] [Doctoral dissertation]. Vanderbilt University; 2012. [cited 2021 Jan 19].
Available from: http://hdl.handle.net/1803/13924.
Council of Science Editors:
Lu LX. Phosphoregulation of the Cdc25 phosphatase and its effects on Schizosaccharomyces pombe mitotic entrance and exit. [Doctoral Dissertation]. Vanderbilt University; 2012. Available from: http://hdl.handle.net/1803/13924
Vanderbilt University
2.
Coffa, Sergio.
Non-visual arrestins bind mitogen activated protein kinases and regulate their signaling.
Degree: PhD, Pharmacology, 2011, Vanderbilt University
URL: http://hdl.handle.net/1803/13814
► Arrestins are multifunctional signaling proteins, important for the regulation of signal transduction and the trafficking of G protein-coupled receptors (GPCRs). Recently, GPCR-arrestin interactions have been…
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▼ Arrestins are multifunctional signaling proteins, important for the regulation of signal transduction and the trafficking of G protein-coupled receptors (GPCRs). Recently, GPCR-arrestin interactions have been proposed to be necessary for activation of G-protein-independent signaling pathways, one of which is the activation of mitogen activated protein kinases (MAPKs). To investigate potential arrestin-MAPK interactions, we have used a variety of molecular tools including the co-expression of the individual domains of arrestin with single components of the c-Raf1-MEK1-ERK2 signaling cascade. We found that non-visual arrestins bind all three kinases, assembling c-Raf1, MEK1, and ERK2 along their short axis, with each kinase directly interacting with both domains.
To further investigate the interactions between arrestins and MAPK, we used alanine-scanning mutagenesis of residues on the non-receptor-binding surface of arrestin that are conserved between arrestin-2 and arrestin-3. We found that the substitution of arginine 307 with an alanine significantly reduced arrestin-2 binding to c-Raf1, whereas the interactions of this mutant with active phosphorylated receptors and the downstream kinases MEK1 and ERK2 were not affected. In contrast to wild type arrestin-2, Arg307Ala mutant failed to rescue arrestin-dependent ERK1/2 activation in arrestin-2/3 knockout MEFs. Interestingly, alanine substitution of the homologous arrestin-3 residue (lysine 308) did not significantly affect c-Raf1 binding or its ability to promote ERK1/2 activation. Together, these findings suggest that the two non-visual arrestins perform the same function via distinct molecular mechanisms. To further elucidate arrestin-MAPK interactions, we performed in vitro binding assays using pure proteins, and demonstrated that ERK2 directly binds free arrestin-2 and arrestin-3, as well as receptor-associated arrestin-1, arrestin-2, and arrestin-3. We have also shown that the arrestin-2 and arrestin-3 association with beta2-adrenergic receptors (β2ARs) significantly enhances ERK2 binding, yet has virtually no effect upon arrestins interactions with the upstream kinases c-Raf1 and MEK1.
Arrestins exist in three conformational states: free, receptor-bound, and microtubule (MT)-bound. Using conformationally biased arrestin mutants, we found that ERK2 prefers two conformations: MT-bound, mimicked by “constitutively inactive” arrestin-Δ7, and receptor-bound, mimicked by “pre-activated” arrestin-3A. Both mutants were able to rescue arrestin-mediated ERK1/2 activation in arrestin-2/3 double knockout fibroblasts. Lastly, we found that the arrestin-2 interaction with c-Raf1 is enhanced by receptor binding, whereas the interaction between arrestin-3 and c-Raf1 is not, thus suggesting that the two non-visual arrestins execute similar functions via diverse mechanisms.
Advisors/Committee Members: Charles Sanders (committee member), Brian E. Wadzinski (committee member), H. Alex Brown (committee member), Vsevolod V. Gurevich (committee member), Benjamin Spiller (Committee Chair).
Subjects/Keywords: arrestin
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APA (6th Edition):
Coffa, S. (2011). Non-visual arrestins bind mitogen activated protein kinases and regulate their signaling. (Doctoral Dissertation). Vanderbilt University. Retrieved from http://hdl.handle.net/1803/13814
Chicago Manual of Style (16th Edition):
Coffa, Sergio. “Non-visual arrestins bind mitogen activated protein kinases and regulate their signaling.” 2011. Doctoral Dissertation, Vanderbilt University. Accessed January 19, 2021.
http://hdl.handle.net/1803/13814.
MLA Handbook (7th Edition):
Coffa, Sergio. “Non-visual arrestins bind mitogen activated protein kinases and regulate their signaling.” 2011. Web. 19 Jan 2021.
Vancouver:
Coffa S. Non-visual arrestins bind mitogen activated protein kinases and regulate their signaling. [Internet] [Doctoral dissertation]. Vanderbilt University; 2011. [cited 2021 Jan 19].
Available from: http://hdl.handle.net/1803/13814.
Council of Science Editors:
Coffa S. Non-visual arrestins bind mitogen activated protein kinases and regulate their signaling. [Doctoral Dissertation]. Vanderbilt University; 2011. Available from: http://hdl.handle.net/1803/13814
Vanderbilt University
3.
Bruntz, Ronald Chase.
Insights into the Molecular Mechanisms of Phospholipase D-Mediated Cancer Cell Survival.
Degree: PhD, Pharmacology, 2014, Vanderbilt University
URL: http://hdl.handle.net/1803/10427
► The production of bioactive lipids by phospholipases has long been appreciated as an important mode of cellular communication. Phospholipase D (PLD) enzymes hydrolyze phosphatidylcholine to…
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▼ The production of bioactive lipids by phospholipases has long been appreciated as an important mode of cellular communication. Phospholipase D (PLD) enzymes hydrolyze phosphatidylcholine to generate a choline headgroup and the important lipid second messenger, phosphatidic acid (PtdOH). PLD family members are found in a diverse range of species from viruses to humans and regulate many important physiological processes including cytoskeletal rearrangements, cell migration, immune response, and cell proliferation. As such, PLD promotes oncogenic processes and elevated PLD activity has been documented in many types of cancerous tissue and derived cell lines. PLD activity is associated with cell cycle progression, resistance to apoptotic stimuli, and tumor cell invasion, but the molecular mechanisms of these PLD-mediated processes are largely uncharacterized. The goal of this project was to identify and characterize novel PLD-protein complexes in order to further understand the mechanisms by which PLD promotes cancer growth and survival. In this dissertation, PLD-derived PtdOH is demonstrated to be a novel regulator of pro-survival Akt kinase in glioblastoma cells by regulating membrane recruitment and activation of Akt. Inhibition of PLD enzymatic activity and subsequent Akt activation decreases GBM cell viability by specifically inhibiting autophagic flux. Additionally, PLD is shown to interact with a number of metabolic enzymes and a potential role for the regulation of cellular bioenergetics in GBM is explored. The results of this research provide mechanistic insight into PLD-mediated cancer cell survival.
Advisors/Committee Members: Brian E. Wadzinski (committee member), Daniel C. Liebler (committee member), H. Alex Brown (committee member), Heidi E. Hamm (committee member), Kevin C. Ess (committee member), John H. Exton (Committee Chair).
Subjects/Keywords: phospholipase D; phosphatidic acid; cancer; Akt; cell signaling; autophagy
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APA (6th Edition):
Bruntz, R. C. (2014). Insights into the Molecular Mechanisms of Phospholipase D-Mediated Cancer Cell Survival. (Doctoral Dissertation). Vanderbilt University. Retrieved from http://hdl.handle.net/1803/10427
Chicago Manual of Style (16th Edition):
Bruntz, Ronald Chase. “Insights into the Molecular Mechanisms of Phospholipase D-Mediated Cancer Cell Survival.” 2014. Doctoral Dissertation, Vanderbilt University. Accessed January 19, 2021.
http://hdl.handle.net/1803/10427.
MLA Handbook (7th Edition):
Bruntz, Ronald Chase. “Insights into the Molecular Mechanisms of Phospholipase D-Mediated Cancer Cell Survival.” 2014. Web. 19 Jan 2021.
Vancouver:
Bruntz RC. Insights into the Molecular Mechanisms of Phospholipase D-Mediated Cancer Cell Survival. [Internet] [Doctoral dissertation]. Vanderbilt University; 2014. [cited 2021 Jan 19].
Available from: http://hdl.handle.net/1803/10427.
Council of Science Editors:
Bruntz RC. Insights into the Molecular Mechanisms of Phospholipase D-Mediated Cancer Cell Survival. [Doctoral Dissertation]. Vanderbilt University; 2014. Available from: http://hdl.handle.net/1803/10427
Vanderbilt University
4.
Cleghorn, Whitney Marie.
Arrestins regulate cell spreading and motility via focal adhesion dynamics.
Degree: PhD, Pharmacology, 2012, Vanderbilt University
URL: http://hdl.handle.net/1803/12660
► Arrestins bind G protein-coupled receptors and more than 100 non-receptor partners, regulating various signaling pathways and cellular functions. The interactions of many proteins (e.g., Src,…
(more)
▼ Arrestins bind G protein-coupled receptors and more than 100 non-receptor partners, regulating various signaling pathways and cellular functions. The interactions of many proteins (
e.g., Src, JNK3, ERK½, Mdm2, etc.) with receptor-bound arrestin localize these molecules to receptor-rich membranes. Our recent finding that arrestins bind microtubules and recruit signaling proteins to the cytoskeleton prompted us to investigate whether arrestins affect cell motility and morphology. Here we describe a novel function of arrestins, their direct effect on focal adhesion dynamics. We demonstrate excessive spreading of cells lacking both non-visual arrestins, which is substrate-independent, evident on both fibronectin and poly-D-lysine. Reduced activity of small GTPases RhoA and Rac1 in arrestin-deficient cells is only partially responsible for the cell spreading phenotype. Increased adhesion, reflected by elevated activity of focal adhesion proteins paxillin and focal adhesion kinase, underlies the exaggerated spreading of arrestin-null cells and their reduced motility. The absence of arrestins greatly increases the size and lifespan of focal adhesions, indicating that arrestins are necessary for rapid focal adhesion turnover. Focal adhesions in arrestin-deficient cells are insensitive to microtubules, suggesting that arrestins likely mediate the induction of focal adhesion disassembly upon microtubule regrowth. Overexpression of WT arrestins and their receptor binding-deficient mutants in arrestin-null cells rescues the phenotype, demonstrating that regulation of focal adhesion dynamics by arrestins is receptor-independent. This is the first demonstration that arrestins play a direct role in focal adhesion dynamics.
Advisors/Committee Members: Roy Zent (committee member), Heidi E. Hamm (committee member), Vsevolod V. Gurevich (committee member), Alissa M. Weaver (committee member), Brian E. Wadzinski (Committee Chair).
Subjects/Keywords: structural biology; pharmacology; morphology; cell biology; GPCR; arrestin
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APA (6th Edition):
Cleghorn, W. M. (2012). Arrestins regulate cell spreading and motility via focal adhesion dynamics. (Doctoral Dissertation). Vanderbilt University. Retrieved from http://hdl.handle.net/1803/12660
Chicago Manual of Style (16th Edition):
Cleghorn, Whitney Marie. “Arrestins regulate cell spreading and motility via focal adhesion dynamics.” 2012. Doctoral Dissertation, Vanderbilt University. Accessed January 19, 2021.
http://hdl.handle.net/1803/12660.
MLA Handbook (7th Edition):
Cleghorn, Whitney Marie. “Arrestins regulate cell spreading and motility via focal adhesion dynamics.” 2012. Web. 19 Jan 2021.
Vancouver:
Cleghorn WM. Arrestins regulate cell spreading and motility via focal adhesion dynamics. [Internet] [Doctoral dissertation]. Vanderbilt University; 2012. [cited 2021 Jan 19].
Available from: http://hdl.handle.net/1803/12660.
Council of Science Editors:
Cleghorn WM. Arrestins regulate cell spreading and motility via focal adhesion dynamics. [Doctoral Dissertation]. Vanderbilt University; 2012. Available from: http://hdl.handle.net/1803/12660
Vanderbilt University
5.
Bartlett, Christina Swan.
Role of the prostaglandin E2 receptor EP1 in hypertensive end-organ damage.
Degree: PhD, Pharmacology, 2012, Vanderbilt University
URL: http://hdl.handle.net/1803/14916
► Hypertension is a prevalent disease affecting one in three adults in the United States. Approximately 25 % of the adult population is either not receiving…
(more)
▼ Hypertension is a prevalent disease affecting one in three adults in the United States. Approximately 25 % of the adult population is either not receiving therapy for their hypertension or is unable to control their blood pressure with current therapies, making treatment of hypertension an important public health goal. In blood pressure regulation, PGE2 can act in a pro-hypertensive or anti-hypertensive manner. It has been demonstrated that EP2 and EP4 receptors mediate the vasodepressor actions of PGE2 and EP1 and EP3 receptors mediate the vasopressor actions of PGE2. Additionally, PGE2 and the EP1 receptor have been demonstrated to mediate at least part of the actions of angiotensin II.
I sought to determine the contribution of EP1 and/or EP3 receptors to hypertensive end-organ damage and diabetic nephropathy. In this dissertation, I utilize mice with genetic disruption of EP1 or EP3 receptors and characterize the outcomes of several models of hypertensive organ damage. In the Nphx/DOCA-NaCl/Ang II model of hypertension, I have demonstrated that disruption of EP1 or EP3 can afford substantial protection from end-organ damage and reduce incidence of mortality. The beneficial effects of EP1 disruption, and likely EP3 disruption, appear to be a result of reduction in MAP in this model. The use of another model involving uninephrectomy and Ang II on a 129S6 background suggests the EP1 receptor plays an important role in hypertensive renal disease independent of blood pressure reduction. Furthermore, genetic disruption of EP1 protected eNOS-/- mice from diabetes-induced proteinuria, independent of blood pressure reduction.
In summary, the data presented in this dissertation advances our knowledge of the role of EP1 and EP3 receptors in hypertension and subsequent sequalae and demonstrate a detrimental role of EP1 in this disease. Targeting the EP1 receptor may be a viable pharmaceutical treatment strategy for hypertension and subsequent organ damage.
Advisors/Committee Members: Richard M. Breyer (committee member), Vsevolod V. Gurevich (committee member), Ambra Pozzi (committee member), Alfred L. George, Jr. (committee member), Brian E. Wadzinski (Committee Chair).
Subjects/Keywords: Prostaglandin; Receptor; Hypertension; Diabetes
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APA (6th Edition):
Bartlett, C. S. (2012). Role of the prostaglandin E2 receptor EP1 in hypertensive end-organ damage. (Doctoral Dissertation). Vanderbilt University. Retrieved from http://hdl.handle.net/1803/14916
Chicago Manual of Style (16th Edition):
Bartlett, Christina Swan. “Role of the prostaglandin E2 receptor EP1 in hypertensive end-organ damage.” 2012. Doctoral Dissertation, Vanderbilt University. Accessed January 19, 2021.
http://hdl.handle.net/1803/14916.
MLA Handbook (7th Edition):
Bartlett, Christina Swan. “Role of the prostaglandin E2 receptor EP1 in hypertensive end-organ damage.” 2012. Web. 19 Jan 2021.
Vancouver:
Bartlett CS. Role of the prostaglandin E2 receptor EP1 in hypertensive end-organ damage. [Internet] [Doctoral dissertation]. Vanderbilt University; 2012. [cited 2021 Jan 19].
Available from: http://hdl.handle.net/1803/14916.
Council of Science Editors:
Bartlett CS. Role of the prostaglandin E2 receptor EP1 in hypertensive end-organ damage. [Doctoral Dissertation]. Vanderbilt University; 2012. Available from: http://hdl.handle.net/1803/14916
Vanderbilt University
6.
Strayhorn, William David.
Expression of Dub-1 and Dub-2 and analysis of a role for deubiquitination in the regulation of nuclear factor-kappa B.
Degree: PhD, Cell and Developmental Biology, 2004, Vanderbilt University
URL: http://hdl.handle.net/1803/11876
► The ubiquitin-proteasome pathway (UPP) is the principal mechanism for the selective degradation of short-lived proteins. The proximal signal for UPP-mediated proteolysis is the covalent modification…
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▼ The ubiquitin-proteasome pathway (UPP) is the principal mechanism for the selective degradation of short-lived proteins. The proximal signal for UPP-mediated proteolysis is the covalent modification of target proteins with multiple ubiquitin polypeptides. Protein-ubiquitin conjugation is catalyzed by ubiquitinating enzymes, which assemble the polyubiquitin degradation signal on a target protein. It is postulated that removal of ubiquitin by deubiquitinating enzymes may also regulate protein targeting to the UPP. However, in comparison to ubiquitinating enzymes, relatively little is known about the functions or regulation of deubiquitinating enzymes. Dub-1 and Dub-2 are closely related deubiquitinating enzymes that were initially identified in hematopoietic cell lines as cytokine-inducible proteins. To gain insights into the substrate(s) and function(s) of the Dub enzymes, I examined the expression of Dub-1 and Dub-2 mRNA and protein, and investigated a potential role for these enzymes in regulation of signal transduction through the nuclear factor-kappa B (NF-kB) family of transcription factors. Expression analyses indicated that Dub-1 is expressed in the developing murine limb bud and in interleukin-3-stimulated FL5.12 pro-B cells. To investigate whether the principal inhibitory protein of NF-kB, IkBa, is a substrate for Dub-1 and Dub-2, a novel in vitro deubiquitination assay was established using polyubiquitinated IkBa as the substrate. In addition, I provide evidence for an IkBa-directed deubiquitinating activity in cytoplasmic lysates from a panel of cell lines. Using this and other complementary assays, I show that Dub-1 and Dub-2 do not deubiquitinate IkBa, do not stabilize IkBa, and do not modulate NF-kB activity. In addition, I show that Dub-1, but not the closely homologous Dub-2, is degraded via the UPP in HEK-293T cells. The UPP-mediated degradation of Dub-1 does not require an intact Dub-1 catalytic domain, thus indicating that this process does not proceed via Dub-1-catalyzed transfer of ubiquitin from a substrate to itself. Overall, these studies provide valuable insights as to the regulation of Dub-1 and Dub-2 that may help elucidate the substrate(s) and biological role(s) of these enzymes. Furthermore, the reagents generated for this dissertation will be useful for the study of Dub biochemistry and IkBa deubiquitination.
Advisors/Committee Members: Virginia L. Shepherd (committee member), Stephen R. Hann (committee member), Eugene M. Oltz (committee member), Brian E. Wadzinski (Committee Chair), David Greenstein (Committee Chair).
Subjects/Keywords: I kappa B alpha; nuclear factor-kappa B; UBP; deubiquitination; DUB; ubiquitin
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APA (6th Edition):
Strayhorn, W. D. (2004). Expression of Dub-1 and Dub-2 and analysis of a role for deubiquitination in the regulation of nuclear factor-kappa B. (Doctoral Dissertation). Vanderbilt University. Retrieved from http://hdl.handle.net/1803/11876
Chicago Manual of Style (16th Edition):
Strayhorn, William David. “Expression of Dub-1 and Dub-2 and analysis of a role for deubiquitination in the regulation of nuclear factor-kappa B.” 2004. Doctoral Dissertation, Vanderbilt University. Accessed January 19, 2021.
http://hdl.handle.net/1803/11876.
MLA Handbook (7th Edition):
Strayhorn, William David. “Expression of Dub-1 and Dub-2 and analysis of a role for deubiquitination in the regulation of nuclear factor-kappa B.” 2004. Web. 19 Jan 2021.
Vancouver:
Strayhorn WD. Expression of Dub-1 and Dub-2 and analysis of a role for deubiquitination in the regulation of nuclear factor-kappa B. [Internet] [Doctoral dissertation]. Vanderbilt University; 2004. [cited 2021 Jan 19].
Available from: http://hdl.handle.net/1803/11876.
Council of Science Editors:
Strayhorn WD. Expression of Dub-1 and Dub-2 and analysis of a role for deubiquitination in the regulation of nuclear factor-kappa B. [Doctoral Dissertation]. Vanderbilt University; 2004. Available from: http://hdl.handle.net/1803/11876
Vanderbilt University
7.
Li, Chunsheng.
MyD88: central relay station of Interleukin 1 signaling pathway.
Degree: PhD, Microbiology and Immunology, 2005, Vanderbilt University
URL: http://hdl.handle.net/1803/15268
► As inflammation is a major mechanism of disease, we investigated the signal transduction processes induced by the key inflammatory cytokine Interleukin (IL) 1 beta as…
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▼ As inflammation is a major mechanism of disease, we investigated the signal transduction processes induced by the key inflammatory cytokine Interleukin (IL) 1 beta as well as the genome responses to pathogen-derived proinflammatory agonists. Myeloid differentiation primary response gene (MyD88) is the essential adaptor protein that transduces intracellular signals generated by the IL-1â receptor and multiple Toll-like receptors (TLRs) that recognize diverse pathogen surfaces. The IL-1â receptor complex interacts with MyD88 via the Toll/IL-1 receptor (TIR) domain. Here we identified the MyD88 TIR domain binding sites involved in IL-1â-induced protein-protein interactions. The MyD88 TIR domain required Box3 to act as dominant negative inhibitor of IL-1â signaling. Accordingly, mutations of residues 285-286 reversed the dominant negative effect of the MyD88 TIR domain on NFêB-dependent reporter gene activity and IL-6 production. Moreover, mutations of residues 171 in helix áA, 195-197 in Box2, and 275 in the â
E strand had similar functional effects. Strikingly, mutations of residues 195-197 eliminated the TIR-TIR interaction of MyD88 and IL-1 receptor accessory protein (IL1RAcP). Mutations in Box2 and 3 prevented homotypic MyD88 oligomerization via the TIR domain. Overall, structure-function analysis produced a 3-dimensional docking model of TIR-TIR interaction between MyD88 and IL1RAcP.
Animal models of systemic inflammation induced by staphylococcal enterotoxin B (SEB) and lipopolysaccharide (LPS) were used to study genome-wide transcriptional response. In vivo treatment with SEB induced 134 and 209 genes in spleen cells and T lymphocytes, respectively. Upregulation of these genes was inhibited by blocking NFêB signaling with a cell-penetrating nuclear import inhibitor cSN50 peptide or the IêBáÄN transgene. In vivo treatment of LPS induced upregulation of 1296 genes and downregulation of 1551 genes in the liver and correspondingly, 1109 and 402 genes in the spleen. The genome-wide response to LPS was ablated in TLR-4-deficent C3H/HeJ mice. The cSN50 peptide blocked 547 LPS-inducible and 669 LPS-downregulated genes in the liver, and 105 LPS-inducible and 230 LPS-suppressed genes in the spleen. Thus, nuclear import of NFêB and other stress-responsive transcription factors plays an important role in genome-wide response to microbial inducers of inflammation.
Advisors/Committee Members: Jack J Hawiger (committee member), Brian E Wadzinski (committee member), Dean W. Ballard (committee member), Luc Van Kaer (committee member), Derya Unutmaz (Committee Chair).
Subjects/Keywords: IL-1beta signaling pathway; MyD88; gene expression profiling; inflammation; nuclear import; IL-1 receptor accessary protein; Interleukin-1; Cellular signal transduction
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APA ·
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APA (6th Edition):
Li, C. (2005). MyD88: central relay station of Interleukin 1 signaling pathway. (Doctoral Dissertation). Vanderbilt University. Retrieved from http://hdl.handle.net/1803/15268
Chicago Manual of Style (16th Edition):
Li, Chunsheng. “MyD88: central relay station of Interleukin 1 signaling pathway.” 2005. Doctoral Dissertation, Vanderbilt University. Accessed January 19, 2021.
http://hdl.handle.net/1803/15268.
MLA Handbook (7th Edition):
Li, Chunsheng. “MyD88: central relay station of Interleukin 1 signaling pathway.” 2005. Web. 19 Jan 2021.
Vancouver:
Li C. MyD88: central relay station of Interleukin 1 signaling pathway. [Internet] [Doctoral dissertation]. Vanderbilt University; 2005. [cited 2021 Jan 19].
Available from: http://hdl.handle.net/1803/15268.
Council of Science Editors:
Li C. MyD88: central relay station of Interleukin 1 signaling pathway. [Doctoral Dissertation]. Vanderbilt University; 2005. Available from: http://hdl.handle.net/1803/15268
. 
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