Crystal Structure of the Hemochromatosis Protein HFE and Characterization of Its Interaction with Transferrin Receptor

AbstractHFE is an MHC-related protein that is mutated in the iron-overload disease hereditary hemochromatosis. HFE binds to transferrin receptor (TfR) and reduces its affinity for iron-loaded transferrin, implicating HFE in iron metabolism. The 2.6 Å crystal structure of HFE reveals the locations of hemochromatosis mutations and a patch of histidines that could be involved in pH-dependent interactions. We also demonstrate that soluble TfR and HFE bind tightly at the basic pH of the cell surface, but not at the acidic pH of intracellular vesicles. TfR:HFE stoichiometry (2:1) differs from TfR:transferrin stoichiometry (2:2), implying a different mode of binding for HFE and transferrin to TfR, consistent with our demonstration that HFE, transferrin, and TfR form a ternary complex

Nuclear factors involved in mitochondrial translation cause a subgroup of combined respiratory chain deficiency.

Mutations in several mitochondrial DNA and nuclear genes involved in mitochondrial protein synthesis have recently been reported in combined respiratory chain deficiency, indicating a generalized defect in mitochondrial translation. However, the number of patients with pathogenic mutations is small, implying that nuclear defects of mitochondrial translation are either underdiagnosed or intrauterine lethal. No comprehensive studies have been reported on large cohorts of patients with combined respiratory chain deficiency addressing the role of nuclear genes affecting mitochondrial protein synthesis to date. We investigated a cohort of 52 patients with combined respiratory chain deficiency without causative mitochondrial DNA mutations, rearrangements or depletion, to determine whether a defect in mitochondrial translation defines the pathomechanism of their clinical disease. We followed a combined approach of sequencing known nuclear genes involved in mitochondrial protein synthesis (EFG1, EFTu, EFTs, MRPS16, TRMU), as well as performing in vitro functional studies in 22 patient cell lines. The majority of our patients were children (<15 years), with an early onset of symptoms <1 year of age (65%). The most frequent clinical presentation was mitochondrial encephalomyopathy (63%); however, a number of patients showed cardiomyopathy (33%), isolated myopathy (15%) or hepatopathy (13%). Genomic sequencing revealed compound heterozygous mutations in the mitochondrial transfer ribonucleic acid modifying factor (TRMU) in a single patient only, presenting with early onset, reversible liver disease. No pathogenic mutation was detected in any of the remaining 51 patients in the other genes analysed. In vivo labelling of mitochondrial polypeptides in 22 patient cell lines showed overall (three patients) or selective (four patients) defects of mitochondrial translation. Immunoblotting for mitochondrial proteins revealed decreased steady state levels of proteins in some patients, but normal or increased levels in others, indicating a possible compensatory mechanism. In summary, candidate gene sequencing in this group of patients has a very low detection rate (1/52), although in vivo labelling of mitochondrial translation in 22 patient cell lines indicate that a nuclear defect affecting mitochondrial protein synthesis is responsible for about one-third of combined respiratory chain deficiencies (7/22). In the remaining patients, the impaired respiratory chain activity is most likely the consequence of several different events downstream of mitochondrial translation. Clinical classification of patients with biochemical analysis, genetic testing and, more importantly, in vivo labelling and immunoblotting of mitochondrial proteins show incoherent results, but a systematic review of these data in more patients may reveal underlying mechanisms, and facilitate the identification of novel factors involved in combined respiratory chain deficiency

Digital Signal Processing

Contains an introduction and reports on fifteen research projects.National Science Foundation FellowshipU.S. Navy - Office of Naval Research (Contract N00014-81-K-0742)National Science Foundation (Grant ECS 84-07285)Sanders Associates, Inc.U.S. Air Force - Office of Scientific Research (Contract F19628-85-K-0028)AT&T Bell Laboratories Doctoral Support ProgramCanada, Bell Northern Research ScholarshipCanada, Fonds pour la Formation de Chercheurs et /'Aide a la Recherche Postgraduate FellowshipCanada, Natural Science and Engineering Research Council Postgraduate FellowshipAmoco Foundation FellowshipFannie and John Hertz Foundation Fellowshi

Digital Signal Processing Research Program

Contains table of contents for Part III, table of contents for Section 1, an introduction and reports on seventeen research projects.U.S. Navy - Office of Naval Research Contract N00014-90-J-1544Charles S. Draper Laboratory Contract DL-H-404158Rockwell Corporation Doctoral FellowshipU.S. Navy - Office of Naval Research Grant N00014-89-J-1489U.S. Navy - Office of Naval Research Grant N00014-90-J-1109The Federative Republic of Brazil ScholarshipLockheed Sanders, Inc.National Science Foundation Grant MIP 87-14969AT&T Bell Laboratories Doctoral ProgramBell Northern Research Ltd.Defense Advanced Research Projects Agency Contract N00014-87-K-0825IBM CorporationSloan FoundationU.S. Air Force - Office of Scientific Research FellowshipU.S. Air Force - Office of Scientific Research Grant AFOSR-91-0034National Science Foundation Graduate FellowshipCanada, Natural Science and Engineering Research Council ScholarshipU.S. Air Force - Office of Scientific Research Grant AFOSR-91-0034Texas Instruments, Inc

Identification and validation of G protein-coupled receptors modulating flow-dependent signaling pathways in vascular endothelial cells

Vascular endothelial cells are exposed to mechanical forces due to their presence at the interface between the vessel wall and flowing blood. The patterns of these mechanical forces (laminar vs. turbulent) regulate endothelial cell function and play an important role in determining endothelial phenotype and ultimately cardiovascular health. One of the key transcriptional mediators of the positive effects of laminar flow patterns on endothelial cell phenotype is the zinc-finger transcription factor, krüppel-like factor 2 (KLF2). Given its importance in maintaining a healthy endothelium, we sought to identify endothelial regulators of the KLF2 transcriptional program as potential new therapeutic approaches to treating cardiovascular disease. Using an approach that utilized both bioinformatics and targeted gene knockdown, we identified endothelial GPCRs capable of modulating KLF2 expression. Genetic screening using siRNAs directed to these GPCRs identified 12 potential GPCR targets that could modulate the KLF2 program, including a subset capable of regulating flow-induced KLF2 expression in primary endothelial cells. Among these targets, we describe the ability of several GPCRs (GPR116, SSTR3, GPR101, LGR4) to affect KLF2 transcriptional activation. We also identify these targets as potential validated targets for the development of novel treatments targeting the endothelium. Finally, we highlight the initiation of drug discovery efforts for LGR4 and report the identification of the first known synthetic ligands to this receptor as a proof-of-concept for pathway-directed phenotypic screening to identify novel drug targets

Digital Signal Processing

Contains table of contents for Part III, table of contents for Section 1, an introduction and reports on seventeen research projects.National Science Foundation FellowshipNational Science Foundation (Grant ECS 84-07285)National Science Foundation (Grant MIP 87-14969)U.S. Navy - Office of Naval Research (Contract N00014-81-K-0742)Scholarship from the Federative Republic of BrazilU.S. Air Force - Electronic Systems Division (Contract F19628-85-K-0028)AT&T Bell Laboratories Doctoral Support ProgramCanada, Bell Northern Research ScholarshipCanada, Fonds pour la Formation de Chercheurs et I'Aide a la Recherche Postgraduate FellowshipSanders Associates, Inc.OKI Semiconductor, Inc.Tel Aviv University, Department of Electronic SystemsU.S. Navy - Office of Naval Research (Contract N00014-85-K-0272)Natural Sciences and Engineering Research Council of Canada, Science and Engineering Scholarshi

Digital Signal Processing

Contains an introduction and reports on twenty research projects.National Science Foundation (Grant ECS 84-07285)U.S. Navy - Office of Naval Research (Contract N00014-81-K-0742)National Science Foundation FellowshipSanders Associates, Inc.U.S. Air Force - Office of Scientific Research (Contract F19628-85-K-0028)Canada, Bell Northern Research ScholarshipCanada, Fonds pour la Formation de Chercheurs et l'Aide a la Recherche Postgraduate FellowshipCanada, Natural Science and Engineering Research Council Postgraduate FellowshipU.S. Navy - Office of Naval Research (Contract N00014-81-K-0472)Fanny and John Hertz Foundation FellowshipCenter for Advanced Television StudiesAmoco Foundation FellowshipU.S. Air Force - Office of Scientific Research (Contract F19628-85-K-0028

A Widespread Chromosomal Inversion Polymorphism Contributes to a Major Life-History Transition, Local Adaptation, and Reproductive Isolation

A set of experiments demonstrates the involvement of a chromosomal inversion in the adaptive transition between annual and perennial ecotypes of the yellow monkeyflower, Mimulus guttatu

Signal Processing Research Program

Contains table of contents for Part III, table of contents for Section 1, an introduction and reports on fourteen research projects.Charles S. Draper Laboratory Contract DL-H-404158U.S. Navy - Office of Naval Research Grant N00014-89-J-1489National Science Foundation Grant MIP 87-14969Battelle LaboratoriesTel-Aviv University, Department of Electronic SystemsU.S. Army Research Office Contract DAAL03-86-D-0001The Federative Republic of Brazil ScholarshipSanders Associates, Inc.Bell Northern Research, Ltd.Amoco Foundation FellowshipGeneral Electric FellowshipNational Science Foundation FellowshipU.S. Air Force - Office of Scientific Research FellowshipU.S. Navy - Office of Naval Research Grant N00014-85-K-0272Natural Science and Engineering Research Council of Canada - Science and Technology Scholarshi

The "unnatural" history of colorectal cancer in Lynch syndrome : Lessons from colonoscopy surveillance

Individuals with Lynch syndrome (LS), one of the most common inherited cancer syndromes, are at increased risk of developing malignancies, in particular colorectal cancer (CRC). Regular colonoscopy with polypectomy is recommended to reduce CRC risk in LS individuals. However, recent independent studies demonstrated that a substantial proportion of LS individuals develop CRC despite regular colonoscopy. The reasons for this surprising observation confirmed by large prospective studies are a matter of debate. In this review, we collect existing evidence from clinical, epidemiological and molecular studies and interpret them with regard to the origins and progression of LS-associated CRC. Alongside with hypotheses addressing colonoscopy quality and pace of progression from adenoma to cancer, we discuss the role of alternative precursors and of immune system in LS-associated CRC. We also identify gaps in current knowledge and make suggestions for future studies aiming at improved CRC prevention for LS individuals.Peer reviewe