Maintenance of cell junctions plays a crucial role in the regulation of cellular functions including cell proliferation, permeability, and cell death. Disruption of cell junctions is implicated in a variety of human d...Maintenance of cell junctions plays a crucial role in the regulation of cellular functions including cell proliferation, permeability, and cell death. Disruption of cell junctions is implicated in a variety of human disorders, such as inflammatory diseases and cancers. Understanding molecular regulation of cell junctions is important for development of therapeutic strategies for intervention of human diseases. Ubiquitination is an important type of post-translational modification that primarily regulates endogenous protein stability, recep- tor internalization, enzyme activity, and protein-protein interactions. Ubiquitination is tightly regulated by ubiq- uitin E3 ligases and can be reversed by deubiquitinating enzymes. Recent studies have been focusing on inves- tigating the effect of protein stability in the regulation of cell-cell junctions. Ubiquitination and degradation of cadherins, claudins, and their interacting proteins are implicated in epithelial and endothelial barrier disruption. Recent studies have revealed that ubiquitination is involved in regulation of Rho GTPases＇ biological activities. Taken together these studies, ubiquitination plays a critical role in modulating cell junctions and motility. In this review, we will discuss the effects of ubiquitination and deubiquitination on protein stability and expression of key proteins in the cell-cell junctions, including junction proteins, their interacting proteins, and small Rho GTPases. We provide an overview of protein stability in modulation of epithelial and endothelial barrier integrity and introduce potential future search directions to better understand the effects of ubiquitination on human disorders caused by dysfunction of cell junctions.展开更多
BACKGROUND： Rho GTPase family members have been shown to participate in neurite growth by regulating the neuronal cytoskeleton. However, there are very few reports of developmental roles of signaling molecules relate...BACKGROUND： Rho GTPase family members have been shown to participate in neurite growth by regulating the neuronal cytoskeleton. However, there are very few reports of developmental roles of signaling molecules related to Rho GTPases. OBJECTIVE： To investigate messenger ribonucleic acid mRNA expression of signaling molecules associated with Rho GTPases, including Rho-A, Rac-1, collapsin response mediator protein 1 （CRMP-1）, and tubulin 133 （Tub/33） during rat hippocampus development. DESIGN, TIME AND SETTING- A non-randomized, controlled, animal experiment, based on different developmental stages of the rat hippocampus, was performed at the Guangdong Key Laboratory of Tissue Construction and Detection, Institute of Clinical Anatomy, Southern Medical University between December 2005 and July 2007. MATERIALS： Trizol reagent was purchased from Invitrogen, USA. RNA PCR kit （AMV） Ver 3.0 and 150 bp DNA Ladder Marker were purchased from TaKaRa, Japan. Unless otherwise specified, all other reagents were purchased from Sigma-Aldrich, USA. METHODS： Twenty-five Sprague Dawley rats were assigned to five groups （n = 5） according to developmental stages： embryonic （embryonic 15 days）, neonatal （postnatal 5 days）, juvenile （postnatal 1 month）, adult （postnatal 3 months）, and senile （postnatal 18 months）. MAIN OUTCOME MEASURES： Detection of mRNA expression of Rho-A, Rac-1, CRMP-1, and Tub β3 during various hippocampal developmental stages by reverse-transcription polymerase chain reaction. RESULTS： Hippocampal mRNA expression of Rho-A, as well as Rac-1, reached peak levels at embryonic, juvenile, and senile stages, and was relatively less during neonatal and adult stages. mRNA expression of Rac-1 was greater than Rho-A during each hippocampal developmental stage. CRMP-1 mRNA expression levels were as follows： embryonic 〉 neonatal 〉 juvenile 〉 adult 〈 senile, while Tubβ3 mRNA expression was embryonic 〉 neonatal 〉 juvenile 〉 adult = senile. CONCLUSION： Rho-A and展开更多
As a critical guanine nucleotide exchange factor(GEF) regulating neurite outgrowth, Trio coordinates multiple processes of cytoskeletal dynamics through activating Rac1, Cdc42 and RhoA small GTPases by two GEF domains...As a critical guanine nucleotide exchange factor(GEF) regulating neurite outgrowth, Trio coordinates multiple processes of cytoskeletal dynamics through activating Rac1, Cdc42 and RhoA small GTPases by two GEF domains, but the in vivo roles of these GEF domains and corresponding downstream effectors have not been determined yet. We established multiple lines of knockout mice and assessed the respective roles of Trio GEF domains and Rac1 in axon outgrowth. Knockout of total Trio in cerebellar granule neurons(CGNs) led to an impaired F-actin rearrangement of growth cone and hence a retarded neurite outgrowth. Such a retardation was reproduced by inhibition of GEF1 domain or knockdown of Cdc42 and restored apparently by introduction of active Cdc42. As Rac1 deficiency did not affect the neurite outgrowth of CGNs, we suggested that Trio GEF1-mediated Cdc42 activation was required for neurite outgrowth. We established a GEF2-knockout line with deletion of all Trio isoforms except a cerebella-specific Trio8, a short isoform of Trio without GEF2 domain, and used this line as a GEF2-deficient animal model. The GEF2-deficient CGNs had a normal neurite outgrowth but abolished Netrin-1-promoted growth, without affecting Netrin-1 induced Rac1 activation. We thus suggested that Trio GEF1-mediated Cdc42 activation rather than Rac1 activation drives the F-actin dynamics necessary for neurite outgrowth, while GEF2 functions in Netrin-1-promoted neurite elongation. Our results delineated the distinct roles of Trio GEF domains in neurite outgrowth, which is instructive to understand the pathogenesis of clinical Trio-related neurodevelopmental disorders.展开更多
文摘Maintenance of cell junctions plays a crucial role in the regulation of cellular functions including cell proliferation, permeability, and cell death. Disruption of cell junctions is implicated in a variety of human disorders, such as inflammatory diseases and cancers. Understanding molecular regulation of cell junctions is important for development of therapeutic strategies for intervention of human diseases. Ubiquitination is an important type of post-translational modification that primarily regulates endogenous protein stability, recep- tor internalization, enzyme activity, and protein-protein interactions. Ubiquitination is tightly regulated by ubiq- uitin E3 ligases and can be reversed by deubiquitinating enzymes. Recent studies have been focusing on inves- tigating the effect of protein stability in the regulation of cell-cell junctions. Ubiquitination and degradation of cadherins, claudins, and their interacting proteins are implicated in epithelial and endothelial barrier disruption. Recent studies have revealed that ubiquitination is involved in regulation of Rho GTPases＇ biological activities. Taken together these studies, ubiquitination plays a critical role in modulating cell junctions and motility. In this review, we will discuss the effects of ubiquitination and deubiquitination on protein stability and expression of key proteins in the cell-cell junctions, including junction proteins, their interacting proteins, and small Rho GTPases. We provide an overview of protein stability in modulation of epithelial and endothelial barrier integrity and introduce potential future search directions to better understand the effects of ubiquitination on human disorders caused by dysfunction of cell junctions.
基金Supported by:the National Basic Research Program of China(973 Program),No. 2007CB512705the Natural Science Foundation of Guangdong Province,No. 8451063201000193
文摘BACKGROUND： Rho GTPase family members have been shown to participate in neurite growth by regulating the neuronal cytoskeleton. However, there are very few reports of developmental roles of signaling molecules related to Rho GTPases. OBJECTIVE： To investigate messenger ribonucleic acid mRNA expression of signaling molecules associated with Rho GTPases, including Rho-A, Rac-1, collapsin response mediator protein 1 （CRMP-1）, and tubulin 133 （Tub/33） during rat hippocampus development. DESIGN, TIME AND SETTING- A non-randomized, controlled, animal experiment, based on different developmental stages of the rat hippocampus, was performed at the Guangdong Key Laboratory of Tissue Construction and Detection, Institute of Clinical Anatomy, Southern Medical University between December 2005 and July 2007. MATERIALS： Trizol reagent was purchased from Invitrogen, USA. RNA PCR kit （AMV） Ver 3.0 and 150 bp DNA Ladder Marker were purchased from TaKaRa, Japan. Unless otherwise specified, all other reagents were purchased from Sigma-Aldrich, USA. METHODS： Twenty-five Sprague Dawley rats were assigned to five groups （n = 5） according to developmental stages： embryonic （embryonic 15 days）, neonatal （postnatal 5 days）, juvenile （postnatal 1 month）, adult （postnatal 3 months）, and senile （postnatal 18 months）. MAIN OUTCOME MEASURES： Detection of mRNA expression of Rho-A, Rac-1, CRMP-1, and Tub β3 during various hippocampal developmental stages by reverse-transcription polymerase chain reaction. RESULTS： Hippocampal mRNA expression of Rho-A, as well as Rac-1, reached peak levels at embryonic, juvenile, and senile stages, and was relatively less during neonatal and adult stages. mRNA expression of Rac-1 was greater than Rho-A during each hippocampal developmental stage. CRMP-1 mRNA expression levels were as follows： embryonic 〉 neonatal 〉 juvenile 〉 adult 〈 senile, while Tubβ3 mRNA expression was embryonic 〉 neonatal 〉 juvenile 〉 adult = senile. CONCLUSION： Rho-A and
基金supported by grants from the National Natural Science Foundation of China (31272311 and 31330034) to M.S.Z.
文摘As a critical guanine nucleotide exchange factor(GEF) regulating neurite outgrowth, Trio coordinates multiple processes of cytoskeletal dynamics through activating Rac1, Cdc42 and RhoA small GTPases by two GEF domains, but the in vivo roles of these GEF domains and corresponding downstream effectors have not been determined yet. We established multiple lines of knockout mice and assessed the respective roles of Trio GEF domains and Rac1 in axon outgrowth. Knockout of total Trio in cerebellar granule neurons(CGNs) led to an impaired F-actin rearrangement of growth cone and hence a retarded neurite outgrowth. Such a retardation was reproduced by inhibition of GEF1 domain or knockdown of Cdc42 and restored apparently by introduction of active Cdc42. As Rac1 deficiency did not affect the neurite outgrowth of CGNs, we suggested that Trio GEF1-mediated Cdc42 activation was required for neurite outgrowth. We established a GEF2-knockout line with deletion of all Trio isoforms except a cerebella-specific Trio8, a short isoform of Trio without GEF2 domain, and used this line as a GEF2-deficient animal model. The GEF2-deficient CGNs had a normal neurite outgrowth but abolished Netrin-1-promoted growth, without affecting Netrin-1 induced Rac1 activation. We thus suggested that Trio GEF1-mediated Cdc42 activation rather than Rac1 activation drives the F-actin dynamics necessary for neurite outgrowth, while GEF2 functions in Netrin-1-promoted neurite elongation. Our results delineated the distinct roles of Trio GEF domains in neurite outgrowth, which is instructive to understand the pathogenesis of clinical Trio-related neurodevelopmental disorders.
