scholarly journals Genetic and biochemical analysis of the adenylyl cyclase of Schizosaccharomyces pombe.

1991 ◽  
Vol 2 (2) ◽  
pp. 155-164 ◽  
Author(s):  
M Kawamukai ◽  
K Ferguson ◽  
M Wigler ◽  
D Young
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Adenylyl Cyclase ◽  
Regulatory Protein ◽  
Wild Type ◽  
Amino Terminal ◽  
Terminal Domain ◽  
Amino Terminal Domain ◽  
Camp Levels ◽  
Adh1 Promoter

The adenylyl cyclase gene, cyr1, of Schizosaccharomyces pombe has been cloned. We have begun an analysis of the function and regulation of adenylyl cyclase by disrupting this gene and by over-expressing all or parts of this gene in various strains. cyr1- strains are viable and contain no measurable cyclic AMP. They conjugate and sporulate under conditions that normally inhibit wild-type strains. Strains containing the cyr1 coding sequences transcribed from the strong adh1 promoter contain greatly elevated adenylyl cyclase activity, as measured in vitro, but only modestly elevated cAMP levels. Such strains conjugate and sporulate less frequently than wild-type cells upon nutrient limitation. Strains which carry the wild-type cyr1 gene but that also express high levels of the amino terminal domain of adenylyl cyclase behave much like cyr1-strains, suggesting that the amino terminal domain can bind a positive regulator. A protein that copurifies with the adenylyl cyclase of S. pombe cross-reacts to antiserum raised against the S. cerevisiae adenylyl cyclase-associated regulatory protein, CAP.

scholarly journals Amino-Terminal Domain Exchange Redirects Origin-Specific Interactions of Adeno-Associated Virus Rep78 In Vitro

2001 ◽  
Vol 75 (7) ◽  
pp. 3230-3239 ◽  
Author(s):  
Miran Yoon ◽  
Deborah H. Smith ◽  
Peter Ward ◽  
Francisco J. Medrano ◽  
Aneel K. Aggarwal ◽  
...  
Keyword(s):  
Dna Replication ◽  
Catalytic Domain ◽  
Adeno Associated Virus ◽  
Free System ◽  
Site Specific ◽  
Amino Terminal ◽  
Terminal Domain ◽  
Site Specific Integration ◽  
Amino Terminal Domain

ABSTRACT The unique ability of adeno-associated virus type 2 (AAV) to site-specifically integrate its genome into a defined sequence on human chromosome 19 (AAVS1) makes it of particular interest for use in targeted gene delivery. The objective underlying this study is to provide evidence for the feasibility of retargeting site-specific integration into selected loci within the human genome. Current models postulate that AAV DNA integration is initiated through the interactions of the products of a single viral open reading frame,REP, with sequences present in AAVS1 that resemble the minimal origin for AAV DNA replication. Here, we present a cell-free system designed to dissect the Rep functions required to target site-specific integration using functional chimeric Rep proteins derived from AAV Rep78 and Rep1 of the closely related goose parvovirus. We show that amino-terminal domain exchange efficiently redirects the specificity of Rep to the minimal origin of DNA replication. Furthermore, we establish that the amino-terminal 208 amino acids of Rep78/68 constitute a catalytic domain of Rep sufficient to mediate site-specific endonuclease activity.

scholarly journals The amino-terminal domain of pyrrolysyl-tRNA synthetase is dispensable in vitro but required for in vivo activity

FEBS Letters ◽  
2007 ◽  
Vol 581 (17) ◽  
pp. 3197-3203 ◽  
Author(s):  
Stephanie Herring ◽  
Alexandre Ambrogelly ◽  
Sarath Gundllapalli ◽  
Patrick O'Donoghue ◽  
Carla R. Polycarpo ◽  
...  
Keyword(s):  
Trna Synthetase ◽  
Amino Terminal ◽  
Terminal Domain ◽  
Amino Terminal Domain

scholarly journals In vitro binding of LexA repressor to DNA: evidence for the involvement of the amino-terminal domain.

1986 ◽  
Vol 5 (4) ◽  
pp. 793-798 ◽  
Author(s):  
S. Hurstel ◽  
M. Granger-Schnarr ◽  
M. Daune ◽  
M. Schnarr
Keyword(s):  
Amino Terminal ◽  
Dna Evidence ◽  
In Vitro Binding ◽  
Terminal Domain ◽  
Lexa Repressor ◽  
Vitro Binding ◽  
Amino Terminal Domain

Ezrin links syndecan-2 to the cytoskeleton

2000 ◽  
Vol 113 (7) ◽  
pp. 1267-1276 ◽  
Author(s):  
F. Granes ◽  
J.M. Urena ◽  
N. Rocamora ◽  
S. Vilaro
Keyword(s):  
Actin Cytoskeleton ◽  
Protein Complex ◽  
Membrane Receptors ◽  
In Vitro Assays ◽  
Amino Terminal ◽  
Terminal Domain ◽  
Rhoa Gtpase ◽  
Triton X ◽  
Amino Terminal Domain

The syndecan family of heparan sulfate proteoglycans is known to associate with the actin cytoskeleton, possibly transducing signals from the extracellular matrix. In the search for proteins that could mediate the association of syndecan-2 with the actin cytoskeleton we found that ezrin, a protein which links membrane receptors to the cytoskeleton, coimmunoprecipitated with syndecan-2 in COS-1 cells. In vitro assays indicated a direct association between the amino-terminal domain of ezrin and the cytoplasmic domain of syndecan-2. Confocal microscopy showed colocalization of ezrin and syndecan-2 in actin-rich microspikes in COS-1 cells. The syndecan-2/ezrin protein complex was resistant to 0.2% Triton X-100 extraction but the syndecan-2/amino-terminal domain of ezrin complex was not, which indicated that carboxi-terminal domain of ezrin is involved in the cytoskeleton anchorage of this protein complex. Additionally we observed that the activation of rhoA GTPase increased syndecan-2 insolubility in 0.2% Triton X-100 and syndecan-2/ezrin association. Taken together, these results indicate that ezrin connects syndecan-2 to the actin cytoskeleton.

scholarly journals Nipah Virus Sequesters Inactive STAT1 in the Nucleus via a P Gene-Encoded Mechanism

2009 ◽  
Vol 83 (16) ◽  
pp. 7828-7841 ◽  
Author(s):  
Michael J. Ciancanelli ◽  
Valentina A. Volchkova ◽  
Megan L. Shaw ◽  
Viktor E. Volchkov ◽  
Christopher F. Basler
Keyword(s):  
Tyrosine Phosphorylation ◽  
Nipah Virus ◽  
Mutant Virus ◽  
Wild Type ◽  
Amino Terminal ◽  
Terminal Domain ◽  
P Gene ◽  
Infected Cells ◽  
Amino Terminal Domain ◽  
In Cells

ABSTRACT The Nipah virus (NiV) phosphoprotein (P) gene encodes the C, P, V, and W proteins. P, V, and W, have in common an amino-terminal domain sufficient to bind STAT1, inhibiting its interferon (IFN)-induced tyrosine phosphorylation. P is also essential for RNA-dependent RNA polymerase function. C is encoded by an alternate open reading frame (ORF) within the common amino-terminal domain. Mutations within residues 81 to 113 of P impaired its polymerase cofactor function, as assessed by a minireplicon assay, but these mutants retained STAT1 inhibitory function. Mutations within the residue 114 to 140 region were identified that abrogated interaction with and inhibition of STAT1 by P, V, and W without disrupting P polymerase cofactor function. Recombinant NiVs were then generated. A G121E mutation, which abrogated inhibition of STAT1, was introduced into a C protein knockout background (Cko) because the mutation would otherwise also alter the overlapping C ORF. In cell culture, relative to the wild-type virus, the Cko mutation proved attenuating but the G121E mutant virus replicated identically to the Cko virus. In cells infected with the wild-type and Cko viruses, STAT1 was nuclear despite the absence of tyrosine phosphorylation. This latter observation mirrors what has been seen in cells expressing NiV W. In the G121E mutant virus-infected cells, STAT1 was not phosphorylated and was cytoplasmic in the absence of IFN stimulation but became tyrosine phosphorylated and nuclear following IFN addition. These data demonstrate that the gene for NiV P encodes functions that sequester inactive STAT1 in the nucleus, preventing its activation and suggest that the W protein is the dominant inhibitor of STAT1 in NiV-infected cells.

scholarly journals A previously unidentified amino-terminal domain regulates transcriptional activity of wild-type and disease-associated human GLI2

2005 ◽  
Vol 14 (15) ◽  
pp. 2181-2188 ◽  
Author(s):  
Erich Roessler ◽  
Alexandre N. Ermilov ◽  
Dorothy Katherine Grange ◽  
Aiqin Wang ◽  
Marina Grachtchouk ◽  
...  
Keyword(s):  
Transcriptional Activity ◽  
Wild Type ◽  
Amino Terminal ◽  
Terminal Domain ◽  
Amino Terminal Domain

An anti-endotoxin peptide that generates from the amino-terminal domain of complement regulatory protein C1 inhibitor

2007 ◽  
Vol 359 (2) ◽  
pp. 285-291 ◽  
Author(s):  
Haimou Zhang ◽  
Jinan Li ◽  
Robert A. Barrington ◽  
Gang Liang ◽  
Gangjian Qin ◽  
...  
Keyword(s):  
Regulatory Protein ◽  
Complement Regulatory Protein ◽  
C1 Inhibitor ◽  
Amino Terminal ◽  
Terminal Domain ◽  
Amino Terminal Domain

scholarly journals A Novel Point Mutation in the Amino Terminal Domain of the Human Glucocorticoid Receptor (hGR) Gene Enhancing hGR-Mediated Gene Expression

2008 ◽  
Vol 93 (12) ◽  
pp. 4963-4968 ◽  
Author(s):  
Evangelia Charmandari ◽  
Takamasa Ichijo ◽  
William Jubiz ◽  
Smita Baid ◽  
Keith Zachman ◽  
...  
Keyword(s):  
Glucocorticoid Receptor ◽  
Molecular Mechanisms ◽  
Nuclear Translocation ◽  
Wild Type ◽  
Exon 2 ◽  
Mutant Receptor ◽  
Amino Terminal ◽  
Terminal Domain ◽  
The Metabolic Syndrome ◽  
Amino Terminal Domain

Context: Interindividual variations in glucocorticoid sensitivity have been associated with manifestations of cortisol excess or deficiency and may be partly explained by polymorphisms in the human glucocorticoid receptor (hGR) gene. We studied a 43-yr-old female, who presented with manifestations consistent with tissue-selective glucocorticoid hypersensitivity. We detected a novel, single, heterozygous nucleotide (G → C) substitution at position 1201 (exon 2) of the hGR gene, which resulted in aspartic acid to histidine substitution at amino acid position 401 in the amino-terminal domain of the hGRα. We investigated the molecular mechanisms of action of the natural mutant receptor hGRαD401H. Methods-Results: Compared with the wild-type hGRα, the mutant receptor hGRαD401H demonstrated a 2.4-fold increase in its ability to transactivate the glucocorticoid-inducible mouse mammary tumor virus promoter in response to dexamethasone but had similar affinity for the ligand (dissociation constant = 6.2 ± 0.6 vs. 6.1 ± 0.6 nm) and time to nuclear translocation (14.75 ± 0.25 vs. 14.25 ± 1.13 min). The mutant receptor hGRαD401H did not exert a dominant positive or negative effect upon the wild-type receptor, it preserved its ability to bind to glucocorticoid response elements, and displayed a normal interaction with the glucocorticoid receptor-interacting protein 1 coactivator. Conclusions: The mutant receptor hGRαD401H enhances the transcriptional activity of glucocorticoid-responsive genes. The presence of the D401H mutation may predispose subjects to obesity, hypertension, and other manifestations of the metabolic syndrome.

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