A5071228734- RNA and protein synthesis mechanisms
- RNA Research and Splicing
- RNA modifications and cancer
- Viral Infections and Immunology Research
- Viral Infectious Diseases and Gene Expression in Insects
- RNA regulation and disease
- Soybean genetics and cultivation
- Legume Nitrogen Fixing Symbiosis
- Monoclonal and Polyclonal Antibodies Research
- Wnt/β-catenin signaling in development and cancer
- Alzheimer's disease research and treatments
- RNA Interference and Gene Delivery
- Respiratory viral infections research
- Bacterial Genetics and Biotechnology
- Stress Responses and Cortisol
- Virus-based gene therapy research
- Transgenic Plants and Applications
- Herpesvirus Infections and Treatments
- Neuropeptides and Animal Physiology
- Toxin Mechanisms and Immunotoxins
- Viral gastroenteritis research and epidemiology
- Thermal Regulation in Medicine
- Genomics and Chromatin Dynamics
- Protein purification and stability
- Plant nutrient uptake and metabolism
Scripps Research Institute
2006-2018
La Jolla Alcohol Research
2013
University of Oxford
2009
Neurosciences Institute
1997-2005
Rockefeller University
1991-1992
McGill University
1985-1986
This study addresses the properties of a newly identified internal ribosome entry site (IRES) contained within mRNA homeodomain protein Gtx. Sequential deletions 5′ untranslated region (UTR) from either end did not define distinct IRES boundaries; when five nonoverlapping UTR fragments were tested, four had activity. These observations are consistent with other cellular analyses suggesting that some IRESes composed segments (IRES modules) independently and combinatorially contribute to...
The expression of Rbm3, a glycine-rich RNA-binding protein, is enhanced under conditions mild hypothermia, and Rbm3 has been postulated to facilitate protein synthesis at colder temperatures. To investigate this possibility, was overexpressed as c-Myc fusion in mouse neuroblastoma N2a cells. Cells expressing showed 3-fold increase both 37°C 32°C compared with control Although polysome profiles cells the were similar, several differences noted, suggesting that might enhance association 40S...
We have identified and characterized a new glycoprotein in the chicken nervous system using immunological molecular biological methods we examined its tissue distribution. Analysis revealed that this protein is very similar structure to neuron-glia cell adhesion molecule, Ng-CAM, mouse L1. cDNA clones encompassing entire coding sequence of Ng-CAM related called Nr-CAM, been isolated sequenced. A containing one major component Mr 145,000 on SDS-PAGE was purified from brain by lentil lectin...
In neurons, translation of dendritically localized mRNAs is thought to play a role in affecting synaptic efficacy. Inasmuch as components the machinery may be limiting dendrites, we investigated mechanisms by which five initiated. The 5′ leader sequences encoding activity-regulated cytoskeletal protein, α subunit calcium–calmodulin-dependent kinase II, dendrin, microtubule-associated protein 2, and neurogranin (RC3) were evaluated for their ability affect untranslated region monocistronic...
The neuron-glia cell adhesion molecule (Ng-CAM) mediates both neuron-neuron and adhesion; it is detected on SDS-PAGE as a predominant 135-kD glycoprotein, with minor components of 80, 190, 210 kD. We have isolated cDNA clones encoding the entire sequence chicken Ng-CAM. predicted extracellular region includes six immunoglobulin-like domains followed by five fibronectin-type III repeats, structural features that are characteristic several neural CAMs N-CAM superfamily. amino acid Ng-CAM most...
In earlier studies, the neural cell adhesion molecule, N-CAM, was found to inhibit proliferation of rat astrocytes both in vitro and vivo . To identify gene targets involved, we used subtractive hybridization examine changes expression that occur after are exposed N-CAM While mRNA levels for decreased such treatment, mRNAs glutamine synthetase calreticulin increased. Since known be involved glucocorticoid receptor pathways, tested a number steroids their effects on astrocyte proliferation....
Cell-free protein synthesis has emerged as a powerful technology for rapid and efficient production. methods are also amenable to automation such systems have been extensively used high-throughput production screening; however, current fluidic not adequate manufacturing biopharmaceuticals. In this work, we report on the initial development of process end-to-end recombinant biologics. This incorporates bioreactor module that can be with eukaryotic or prokaryotic lysates programmed combined...
Although mild hypothermia generally reduces protein synthesis in mammalian cells, the expression of a small number proteins, including Rbm3, is induced under these conditions. In this study, we identify an <i>Rbm3</i> mRNA with complex 5′ leader sequence containing multiple upstream open reading frames. are potentially inhibitory to translation, monocistronic reporter mRNAs were translated relatively efficiently. addition, when tested intercistronic region dicistronic mRNAs, dramatically...
Significance Translation of hepatitis C virus (HCV) RNA is facilitated by an internal ribosome entry site (IRES) that binds small (40S) ribosomal subunits via protein and 18S (rRNA) interactions. Using rRNA expression system developed in this laboratory, we demonstrate IRES activity requires a 3-nt Watson–Crick base-pairing interaction between the 40S subunits. The specific intermolecular appears to be unique HCV potential therapeutic target Hepatitis disease. In addition, expressing...
β-Secretase [also known as the β-site amyloid precursor protein-cleaving enzyme 1 (BACE1)] is an involved in production of Aβ-amyloid plaques brains patients with Alzheimer's disease. The enhanced this occurs without corresponding changes BACE1 mRNA levels. complex 5′ leader contains three upstream ORFs (uORFs) preceding initiation codon. In study, we investigated how affects translation efficiency a first step understanding Using reporter constructs transfected mammalian cell lines and...
Nr-CAM is a membrane glycoprotein that expressed on neurons. It structurally related to members of the N-CAM superfamily neural cell adhesion molecules having six immunoglobulin-like domains and five fibronectin type III repeats in extracellular region. We have found aggregation chick brain cells was inhibited by anti-Nr-CAM Fab' fragments, indicating can act as molecule. To clarify mode action Nr-CAM, mouse fibroblast line L-M(TK-) (or L cells) transfected with DNA expression construct...
In higher eukaryotes, translation of some mRNAs occurs by internal initiation. It is not known, however, whether this mechanism used to initiate the any yeast mRNAs. report, we identify naturally occurring nucleotide sequences that function as ribosome entry sites (IRESes) within 5′ leader Saccharomyces cerevisiae YAP1 and p150 When tested in untranslated regions monocistronic reporter genes, both enhanced efficiency vegetatively growing cells. Moreover, when intercistronic region...
Numerous eukaryotic mRNAs contain sequences complementary to segments of the 18S and 28S rRNAs. Previous studies raised possibility that these complementarities might allow mRNA–rRNA interactions affect rates translation. In present study, we investigated mRNA encoding mouse Gtx homeodomain protein. This contains within its 5′ untranslated region (UTR) a segment is two regions rRNA, located at nucleotides 701–741 1104–1136. A RNA probe containing this complementarity could be photochemically...
Many eukaryotic mRNAs contain sequences that resemble segments of 28S and 18S rRNAs, these rRNA-like are present in both the sense antisense orientations. Some similar to highly conserved regions whereas others have sequence similarities expansion segments. In particular, four found several hundred different genes, location within various genes is not random. One preferentially located protein coding immediately upstream termination codon a number genes. Northern blot analysis poly(A) + RNA...
The nodulin-23 gene of soybean is one the most abundantly transcribed genes induced during symbiosis with Rhizobium. Using a plasmid (pNod25) from nodule cDNA library, we have isolated nodulin genomic library. Nucleotide sequence analysis and clone indicated that coding region this 669 bp long interrupted by single intron about 53ø bp. deduced protein suggests may signal sequence. 5′-flanking two other genes, nodulin-24 encoding for mambrane polypeptide leghanoglobin (LbC 3 ), were obtained....
Our recent demonstration that many eukaryotic mRNAs contain sequences complementary to rRNA led the hypothesis these might mediate specific interactions between and ribosomes thereby affect translation. In present experiments, ability of bind was investigated by using photochemical cross-linking. RNA probes with perfect complementarity 18S or 28S were shown cross-link specifically corresponding within intact ribosomal subunits. Similar results obtained based on natural mRNA varying degrees...
In eukaryotes, 40S ribosomal subunits move from their recruitment site on the mRNA to initiation codon by an as yet poorly understood process. One postulated mechanism involves shunting, in which completely bypass regions of 5' leader. For some mRNAs, shunting has been shown require various elements, are thought base pair 18S rRNA; however, role pairing not tested directly. earlier studies, we demonstrated that a short element leader Gtx homeodomain functioned rRNA. Using model system assess...
Sequences that control translation of mRNA may play critical roles in regulating protein levels. One such element is the internal ribosome entry site (IRES). We previously showed a 9-nt segment 5′ leader sequence encoding Gtx homeodomain could function as an IRES. To identify other short sequences with similar properties, we designed selection procedure uses retroviral vector to express dicistronic mRNAs enhanced green and cyan fluorescent proteins first second cistrons, respectively....