ERK5 (also known as BMK1), a member of the mitogen-activated protein kinase (MAPK) superfamily, was to be activated strongly by oxidant and osmotic stresses. Here we have found that is epidermal growth factor nerve factor, whose receptors are tyrosine kinases. The activation inhibited expression dominant-negative Ras induced active in PC12 cells, indicating requirement for activation. factor-induced PD98059 U0126 inhibitors, which were previously thought act specifically on classical MAPK...

Regulated synthesis of reactive oxygen species (ROS) by membrane-bound fungal NADPH oxidases (Nox) plays a key role in morphogenesis, growth, and development. Generation the plant symbiotic fungus, Epichloë festucae , requires functional assembly multisubunit complex composed NoxA, regulatory component, NoxR, small GTPase RacA. However, mechanism for activation this at plasma membrane is unknown. We found yeast two-hybrid coimmunoprecipitation assays that E. NoxR interacts with homologs...

Recent studies have shown that<i>Drosophila</i> Dishevelled (Dsh), an essential component of the<i>wingless</i> signal transduction, is also involved in planar polarity signaling through the c-Jun N-terminal kinase (JNK)/stress-activated protein (SAPK) pathway in<i>Drosophila</i>. Here, we show that expression a mouse homolog Dsh (mDvl-1) NIH3T3 cells activates JNK/SAPK, and its activator MKK7. A C-terminal half mDvl-1 which contains DEP domain was sufficient for activation whereas as well...

Interaction between the mammalian cell polarity proteins mInsc (mammalian homologue of Inscuteable) and Leu-Gly-Asn repeat-enriched protein (LGN), as well that their respective Drosophila homologues Inscuteable Partner (Pins), plays crucial roles in mitotic spindle orientation, a process contributing to asymmetric division. Here, we report crystal structure LGN-binding domain (LBD) human complexed with N-terminal tetratricopeptide repeat (TPR) motifs LGN at 2.6-Å resolution. In complex,...

Highlights•mInsc regulates directional movement during neutrophil chemotaxis•Chemoattractants induced Gαi-GDP accumulation at the front of migrating neutrophils•Gαi-GDP interacts with mInsc via LGN/AGS3 to recruit Par-aPKC complex•Both and aPKC are required for control, but not cell motilitySummarySuccessful chemotaxis requires only increased motility also sustained directionality. Here, we show that, receptors coupled Gi family heterotrimeric G proteins, is regulated by mInsc, a mammalian...

The membrane-integrated NADPH oxidases DUOX1 and DUOX2 are recruited to the apical plasma membrane in epithelial cells release hydrogen peroxide, thereby playing crucial roles various functions such as thyroid hormone synthesis host defense. However, it has remained unknown about molecular mechanism for sorting of DUOX2. Here we show that correctly sorted via membrane-spanning DUOX maturation proteins DUOXA1 DUOXA2, respectively, when co-expressed MDCK cells. Impairment N-glycosylation...

ABSTRACT The small GTPases RhoA and Cdc42 their effector proteins play crucial roles in neutrophil chemotaxis. However, endogenous localization regulation of these have remained largely unknown. Here, we show, using a trichloroacetic acid fixation method, that are preferentially accumulated at the F‐actin‐rich leading edge (pseudopod) during chemotaxis human neutrophil‐like PLB‐985 cells response to chemoattractant C5a. Interestingly, enrichment is impaired by knockdown Cdc42, indicating...

Formation of apico-basal polarity in epithelial cells is crucial for both morphogenesis (e.g., cyst formation) and function tight junction development). Atypical protein kinase C (aPKC), complexed with Par6, considered to translocate the apical membrane cell polarization. However, mechanism translocation Par6-aPKC complex has remained largely unknown. Here, we show that WD40 Morg1 (mitogen-activated organizer 1) directly binds Par6 thus facilitates targeting Madin-Darby canine kidney cells....

Most cells in tissues are polarized and usually have two distinct plasma membrane domains-an apical a basolateral membrane, which the result of trafficking proteins lipids. However, mechanism underlying cell polarization is not fully understood. In this study, we investigated involvement synaptotagmin-like protein 2-a (Slp2-a), an effector molecule for small GTPase Rab27, by using Madin-Darby canine kidney II as model cells. The results show that level Slp2-a expression MDCK increases...

The adhesion family of G protein-coupled receptors (GPCRs) is defined by an N-terminal large extracellular region that contains various adhesion-related domains and a highly-conserved GPCR-autoproteolysis-inducing (GAIN) domain, the latter which located immediately before canonical seven-transmembrane domain. These are expressed widely involved in functions including development, angiogenesis, synapse formation, tumorigenesis. GPR125 (ADGRA3), orphan GPCR, has been shown to modulate planar...

The superoxide-producing NADPH oxidase in phagocytes is crucial for host defence; its catalytic core the membrane-integrated protein gp91phox [also known as Nox2 (NADPH 2)], which forms a stable heterodimer with p22phox. Activation of requires membrane translocation three cytosolic proteins p47phox, p67phox and small GTPase Rac. At membrane, these assemble gp91phox-p22phox induce conformational change gp91phox, leading to superoxide production. p47phox translocates membranes using two...

Neutrophils play an essential role via phagocytosis in host defense against microbial infections. However, little is known about molecular mechanisms underlying neutrophils, because of the difficulty genetically manipulating these cells. Here, we provide first comprehensive description phospholipid metabolism during human which have efficiently transfected with cDNAs encoding lipid-probing protein modules. Phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P(2)), F-actin organizer abundant...

Bipolar spindle assembly in mitotic cells is a prerequisite to ensure correct alignment of chromosomes for their segregation each daughter cell; microtubules are tethered at plus ends and focused minus either the two poles. NuMA (nuclear apparatus protein) present solely nucleus interphase cells, but relocalizes during mitosis poles play crucial role via focusing pole. In study we show that kinesin-5 family motor Eg5 protein directly interacts with NuMA, using proteomics approach various...

Proper mitotic spindle orientation requires that astral microtubules are connected to the cell cortex by microtubule-binding protein NuMA, which is recruited from cytoplasm. Cortical recruitment of NuMA at least partially mediated via direct binding adaptor LGN. LGN normally adopts a closed conformation an intramolecular interaction between its N-terminal NuMA-binding domain and C-terminal region contains four GoLoco (GL) motifs, each capable membrane-anchored Gαi subunit heterotrimeric G...

Correct cyst morphogenesis of epithelial cells requires apical–basal polarization, which is partly regulated by mitotic spindle orientation, a process dependent on the heterotrimeric G protein subunit Gαi and its binding LGN. Here, we show that in three‐dimensional culture mammalian Madin–Darby canine kidney (MDCK) cells, Gαi‐activating Ric‐8A crucial for orientation formation normal cysts comprise single layer polarized with their apical surfaces lining an inner lumen. Consistent...

// Makoto Kawamoto 1 , Hideya Onishi Keigo Ozono 2 Akio Yamasaki Akira Imaizumi 1, 3 Sachiko Kamakura 4 Kenji Nakano 5 Yoshinao Oda Hideki Sumimoto and Masafumi Nakamura 6 Department of Cancer Therapy Research, Graduate School Medical Sciences, Kyushu University, Fukuoka, Japan Anatomic Pathology, Shukoukai Inc., Tokyo, Biochemistry, Innovation Center for Redox Navigation, Surgery Oncology, Correspondence to: Onishi, email: ohnishi@surg1.med.kyushu-u.ac.jp Keywords: TrkB, BDNF, gallbladder...

A single epithelial cell embedded in extracellular matrix (ECM) can proliferate to form an apical lumen-harboring cyst, whose formation is a fundamental step organ development. At early two-cell stage after division, the doublet typically displays "inverted" polarity, with and basolateral proteins being located ECM-facing cell-cell-contacting plasma membranes, respectively. Correct cystogenesis requires polarity reorientation, process containing protein endocytosis from ECM-abutting...

Transmembrane glycoproteins, synthesized at the endoplasmic reticulum (ER), generally reach Golgi apparatus in COPII‐coated vesicles en route to cell surface. Here, we show that bona fide nonglycoprotein Nox5, a transmembrane superoxide‐producing NADPH oxidase, is transported surface manner resistant co‐expression of Sar1 (H79G), GTP‐fixed mutant small GTPase Sar1, which blocks COPII vesicle fission from ER. In contrast, (H79G) effectively inhibits ER‐to‐Golgi transport glycoproteins...