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Middle Domain (middle + domain)
Selected AbstractsA hydrophobic segment within the C-terminal domain is essential for both client-binding and dimer formation of the HSP90-family molecular chaperoneFEBS JOURNAL, Issue 1 2003Shin-ichi Yamada The , isoform of human 90-kDa heat shock protein (HSP90,) is composed of three domains: the N-terminal (residues 1,400); middle (residues 401,615) and C-terminal (residues 621,732). The middle domain is simultaneously associated with the N- and C-terminal domains, and the interaction with the latter mediates the dimeric configuration of HSP90. Besides one in the N-terminal domain, an additional client-binding site exists in the C-terminal domain of HSP90. The aim of the present study is to elucidate the regions within the C-terminal domain responsible for the bindings to the middle domain and to a client protein, and to define the relationship between the two functions. A bacterial two-hybrid system revealed that residues 650,697 of HSP90, were essential for the binding to the middle domain. An almost identical region (residues 657,720) was required for the suppression of heat-induced aggregation of citrate synthase, a model client protein. Replacement of either Leu665-Leu666 or Leu671-Leu672 to Ser-Ser within the hydrophobic segment (residues 662,678) of the C-terminal domain caused the loss of bindings to both the middle domain and the client protein. The interaction between the middle and C-terminal domains was also found in human 94-kDa glucose-regulated protein. Moreover, Escherichia coli HtpG, a bacterial HSP90 homologue, formed heterodimeric complexes with HSP90, and the 94-kDa glucose-regulated protein through their middle-C-terminal domains. Taken together, it is concluded that the identical region including the hydrophobic segment of the C-terminal domain is essential for both the client binding and dimer formation of the HSP90-family molecular chaperone and that the dimeric configuration appears to be similar in the HSP90-family proteins. [source] Dynamin 2 mutations associated with human diseases impair clathrin-mediated receptor endocytosis,HUMAN MUTATION, Issue 10 2009Marc Bitoun Abstract Dynamin 2 (DNM2) is a large GTPase involved in the release of nascent vesicles during endocytosis and intracellular membrane trafficking. Distinct DNM2 mutations, affecting the middle domain (MD) and the Pleckstrin homology domain (PH), have been identified in autosomal dominant centronuclear myopathy (CNM) and in the intermediate and axonal forms of the Charcot-Marie-Tooth peripheral neuropathy (CMT). We report here the first CNM mutation (c.1948G>A, p.E650,K) in the DNM2 GTPase effector domain (GED), leading to a slowly progressive moderate myopathy. COS7 cells transfected with DNM2 constructs harboring a disease-associated mutation in MD, PH, or GED show a reduced uptake of transferrin and low-density lipoprotein (LDL) complex, two markers of clathrin-mediated receptor endocytosis. A decrease in clathrin-mediated endocytosis was also identified in skin fibroblasts from one CNM patient. We studied the impact of DNM2 mutant overexpression on epidermal growth factor (EGF)-induced extracellular signal-regulated kinase 1 (ERK1) and ERK2 activation, known to be an endocytosis- and DNM2-dependent process. Activation of ERK1/2 was impaired for all the transfected mutants in COS7 cells, but not in CNM fibroblasts. Our results indicate that impairment of clathrin-mediated endocytosis may play a role in the pathophysiological mechanisms leading to DNM2-related diseases, but the tissue-specific impact of DNM2 mutations in both diseases remains unclear. Hum Mutat 30:1,9, 2009. © 2009 Wiley-Liss, Inc. [source] Morphology and sedimentary systems in the Central Bransfield Basin, Antarctic Peninsula: sedimentary dynamics from shelf to basinBASIN RESEARCH, Issue 3 2009Marga García ABSTRACT A detailed regional characterization of the physiography, morphology and sedimentary systems of the Central Bransfield Basin (CBB) was carried out using swath bathymetry and high- and very high-resolution seismic profiles. The basin margins show continental shelves with numerous glacial troughs, and continental slopes where relatively wide and flat slope platforms represent the middle domain in an atypical physiographic scenario in glaciated margins. Although the CBB is tectonically active, most of the morphologic features are sedimentary in origin, and can be classified into four sedimentary systems: (1) glacial-glaciomarine, composed of erosional surfaces, glacial troughs, furrows and draping sheets; (2) slope-basin, formed by trough mouth fans, slope aprons, the Gebra-Magia instability complex and turbidity systems; (3) seabed fluid outflow system composed of pockmark fields; and (4) contourite, composed of drifts and moats. The sedimentary systems show a clear zonation from shelf to basin and their dynamics reflects the complex interplay among glacial, glaciomarine, marine and oceanographic processes involved in the entire shelf-to-basin sediment distribution. The CBB morphology is primarily controlled by glacial/interglacial cyclicity and physiography and to a lesser extent by tectonics and oceanography. These factors have affected the South Shetland Islands (SSI) and Antarctic Peninsula (AP) margins differently, creating a relatively starved SSI margin and a more constructional AP margin. They have also created two entire sediment-dispersal domains: the shelf-to-slope, which records the glaciation history of the CBB; and the lower slope-to-basin, which records the imprint of local factors. This study provides a ,source-to-sink' sedimentary scheme for glaciated margins, which may be applied to the basin research in other margins, based on the characterization of sedimentary systems, their boundaries and the linkages among them. This approach proves to be adequate for the identification of global and local factors governing the CBB and may therefore be applied to other study areas. [source] Crystallization and preliminary X-ray diffraction analysis of the middle domain of Paip1ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 10 2009Ahmad Seif Kanaan The poly(A)-binding protein (PABP) simultaneously interacts with the poly(A) tail of mRNAs and the scaffolding protein eIF4G to mediate mRNA circularization, resulting in stimulation of protein translation. PABP is regulated by the PABP-interacting protein Paip1. Paip1 is thought to act as a translational activator in 5, cap-dependent translation by interacting with PABP and the initiation factors eIF4A and eIF3. Here, the crystallization and preliminary diffraction analysis of the middle domain of Paip1 (Paip1M), which produces crystals that diffract to a resolution of 2.2,Å, are presented. [source] Structural Models and Binding Site Prediction of the C-terminal Domain of Human Hsp90: A New Target for Anticancer DrugsCHEMICAL BIOLOGY & DRUG DESIGN, Issue 5 2008Miriam Sgobba Heat shock protein 90 is a valuable target for anticancer drugs because of its role in the activation and stabilization of multiple oncogenic signalling proteins. While several compounds inhibit heat shock protein 90 by binding the N-terminal domain, recent studies have proved that the C-terminal domain is important for dimerization of the chaperone and contains an additional binding site for inhibitors. Heat shock protein 90 inhibition achieved with molecules binding to the C-terminal domain provides an additional and novel opportunity to design and develop drugs. Therefore, for the first time, we have investigated the structure and the dynamic behaviour of the C-terminal domain of human heat shock protein 90 with and without the small-middle domain, using homology modelling and molecular dynamics simulations. In addition, secondary structure predictions and peptide folding simulations proved useful to investigate a putative additional ,-helix located between H18 and ,20 of the C-terminal domain. Finally, we used the structural information to infer the location of the binding site located in the C-terminal domain by using a number of computational tools. The predicted pocket is formed by two grooves located between helix H18, the loop downstream of H18 and the loop connecting helices H20 and H21 of each monomer of the C-terminal domain, with only two amino acids contributing from each middle domain. [source] The polypeptide chain release factor eRF1 specifically contacts the s4UGA stop codon located in the A site of eukaryotic ribosomesFEBS JOURNAL, Issue 10 2001Laurent Chavatte It has been shown previously [Brown, C.M. & Tate, W.P. (1994) J. Biol. Chem.269, 33164,33170.] that the polypeptide chain release factor RF2 involved in translation termination in prokaryotes was able to photocrossreact with mini-messenger RNAs containing stop signals in which U was replaced by 4-thiouridine (s4U). Here, using the same strategy we have monitored photocrosslinking to eukaryotic ribosomal components of 14-mer mRNA in the presence of , and 42-mer mRNA in the presence of tRNAAsp (tRNAAsp gene transcript). We show that: (a) both 14-mer and 42-mer mRNAs crossreact with ribosomal RNA and ribosomal proteins. The patterns of the crosslinked ribosomal proteins are similar with both mRNAs and sensitive to ionic conditions; (b) the crosslinking patterns obtained with 42-mer mRNAs show characteristic modification upon addition of tRNAAsp providing evidence for appropriate mRNA phasing onto the ribosome. Similar changes are not detected with the 14-mer pairs; (c) when eukaryotic polypeptide chain release factor 1 (eRF1) is added to the ribosome·tRNAAsp complex it crossreacts with the 42-mer mRNA containing the s4UGA stop codon located in the A site, but not with the s4UCA sense codon; this crosslink involves the N-terminal and middle domains of eRF1 but not the C domain which interacts with eukaryotic polypeptide chain release factor 3 (eRF3); (d) addition of eRF3 has no effect on the yield of eRF1,42-mer mRNA crosslinking and eRF3 does not crossreact with 42-mer mRNA. These experiments delineate the in vitro conditions allowing optimal phasing of mRNA on the eukaryotic ribosome and demonstrate a direct and specific contact of ,core' eRF1 and s4UGA stop codon within the ribosomal A site. [source] NPH4/ARF7 and ARF19 promote leaf expansion and auxin-induced lateral root formationTHE PLANT JOURNAL, Issue 1 2005Jill C. Wilmoth Summary Auxin response factors (ARFs) bind auxin response promoter elements and mediate transcriptional responses to auxin. Five of the 22 ARF genes in Arabidopsis thaliana encode ARFs with glutamine-rich middle domains. Four of these can activate transcription and have been ascribed developmental functions. We show that ARF19, the fifth Q-rich ARF, also activates transcription. Mutations in ARF19 have little effect on their own, but in combination with mutations in NPH4/ARF7, encoding the most closely related ARF, they cause several phenotypes including a drastic decrease in lateral and adventitious root formation and a decrease in leaf cell expansion. These results indicate that auxin induces lateral roots and leaf expansion by activating NPH4/ARF7 and ARF19. Auxin induces the ARF19 gene, and NPH4/ARF7 and ARF19 together are required for expression of one of the arf19 mutant alleles, suggesting that a positive feedback loop regulates leaf expansion and/or lateral root induction. [source] | |||||||||||||