Molecular and genetic characterization of the interactions between the Drosophila stoned-B protein and DAP-160 (intersectin)

The stoned locus of Drosophila produces a dicistronic transcript and encodes two proteins, stoned-A (STNA) and stoned-B (STNB). Both proteins are located at synaptic terminals. The STNB protein contains a domain that has homology with the μ-subunit of the AP (adaptor protein) complex, as well as a number of NPF (Asp-Pro-Phe) motifs known to bind EH (Eps15 homology) domains. Mutations at the stoned locus interact synergistically with mutations at the shibire (dynamin) locus and alter synaptic vesicle endocytosis. The STNB protein has also been shown to interact with synaptic vesicles via synaptogamin-I. We initiated an investigation of the possible interaction of DAP-160 (dynamin-associated protein of 160 kDa), a Drosophila member of the intersectin family, with the STNB protein. We show here that both of the viable stoned alleles interacted with a genetic construct that reduces DAP-160 levels to 25% of normal. One of these stoned alleles contains a substitution resulting in a stop codon in the open reading frame encoding STNB. This allele also shows markedly reduced levels of both DAP-160 and dynamin. As anticipated, the NPF motifs in STNB are found to be high-affinity binding motifs for the EH domains of DAP-160. One of the SH3 (Src homology 3) domains of DAP-160 also interacts with STNB. Finally, we show that immunoprecipitation of STNB from fly head extracts co-precipitates with DAP-160, and we conclude that the interaction of the STNB protein with both synaptotagmin I and DAP-160 may regulate synaptic vesicle recycling by recruiting dynamin to a pre-fission complex.

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Interaction of Grb2 SH3 domain with UVRAG in an Alzheimer’s disease–like scenario

Biochemistry and Cell Biology ◽

10.1139/bcb-2014-0001 ◽

2014 ◽

Vol 92 (3) ◽

pp. 219-225 ◽

Cited By ~ 1

Author(s):

Kasturi Roy ◽

Oishee Chakrabarti ◽

Debashis Mukhopadhyay

Keyword(s):

Alzheimer’S Disease ◽

Alzheimer's Disease ◽

Growth Factor Receptor ◽

Adaptor Protein ◽

Sh3 Domain ◽

Binding Motifs ◽

Src Homology 3 ◽

Src Homology ◽

Cellular Compartmentalization

Growth factor receptor-bound protein 2 (Grb2) is an adaptor protein which participates in trafficking pathways alongside its role in signaling. Proteins important for actin remodeling and cellular compartmentalization contain SRC Homology 3 (SH3) binding motifs that interact with Grb2. While studying the Grb2–amyloid precursor protein (APP) intracellular domain (AICD) interaction in Alzheimer’s disease cell line models, it was seen that Grb2 colocalized to compartments that mature into autophagosomes. The entrapping of AICD in the Grb2 vesicles and its clearance via autophagosomes was a survival contrivance on the part of the cell. Here, we report that Grb2, when in excess, interacts with ultraviolet radiation resistance-associated gene protein (UVRAG) under excess conditions of AICD–Grb2 or Grb2. The N-terminal SH3 domain of Grb2 specifically interacts with UVRAG, unlike the C-terminal SH3 domain. This interaction helps to understand the role of Grb2 in the autophagic maturation of vesicles.

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Faculty Opinions recommendation of The neural cell adhesion molecule promotes maturation of the presynaptic endocytotic machinery by switching synaptic vesicle recycling from adaptor protein 3 (AP-3)- to AP-2-dependent mechanisms.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.718146548.793485703 ◽

2013 ◽

Author(s):

Y Peng Loh ◽

Joshua J Park

Keyword(s):

Cell Adhesion ◽

Synaptic Vesicle ◽

Adhesion Molecule ◽

Cell Adhesion Molecule ◽

Neural Cell Adhesion Molecule ◽

Adaptor Protein ◽

Neural Cell ◽

Synaptic Vesicle Recycling ◽

Vesicle Recycling ◽

Neural Cell Adhesion

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Biochemical and Genetic Characterization of Propionicin T1, a New Bacteriocin from Propionibacterium thoenii

Applied and Environmental Microbiology ◽

10.1128/aem.66.10.4230-4236.2000 ◽

2000 ◽

Vol 66 (10) ◽

pp. 4230-4236 ◽

Cited By ~ 40

Author(s):

Therese Faye ◽

Thor Langsrud ◽

Ingolf F. Nes ◽

Helge Holo

Keyword(s):

Amino Acids ◽

Stop Codon ◽

Sequence Similarity ◽

Propionibacterium Freudenreichii ◽

Terminal Part ◽

Reading Frame ◽

Lactobacillus Sake ◽

Propionibacterium Thoenii ◽

Self Protection

ABSTRACT A collection of propionibacteria was screened for bacteriocin production. A new bacteriocin named propionicin T1 was isolated from two strains of Propionibacterium thoenii. This bacteriocin shows no sequence similarity to other bacteriocins. Propionicin T1 was active against all strains of Propionibacterium acidipropionici, Propionibacterium thoenii, andPropionibacterium jensenii tested and also againstLactobacillus sake NCDO 2714 but showed no activity againstPropionibacterium freudenreichii. The bacteriocin was purified, and the N-terminal part of the peptide was determined with amino acid sequencing. The corresponding gene pctA was sequenced, and this revealed that propionicin T1 is produced as a prebacteriocin of 96 amino acids with a typical sec leader, which is processed to give a mature bacteriocin of 65 amino acids. An open reading frame encoding a protein of 424 amino acids was found 68 nucleotides downstream the stop codon of pctA. The N-terminal part of this putative protein shows strong similarity with the ATP-binding cassette of prokaryotic and eukaryotic ABC transporters, and this protein may be involved in self-protection against propionicin T1. Propionicin T1 is the first bacteriocin from propionibacteria that has been isolated and further characterized at the molecular level.

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Modulation of Src Homology 3 Proteins by the Proline-Rich Adaptor Protein Shb

Experimental Cell Research ◽

10.1006/excr.1996.3471 ◽

1997 ◽

Vol 231 (2) ◽

pp. 269-275 ◽

Cited By ~ 3

Author(s):

Torbjörn Karlsson ◽

Michael Welsh

Keyword(s):

Adaptor Protein ◽

Src Homology 3 ◽

Src Homology

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Novel Src Homology 3 Domain-binding Motifs Identified from Proteomic Screen of a Pro-rich Region

Molecular & Cellular Proteomics ◽

10.1074/mcp.m500108-mcp200 ◽

2005 ◽

Vol 4 (8) ◽

pp. 1155-1166 ◽

Cited By ~ 49

Author(s):

Christina Y. H. Jia ◽

Jing Nie ◽

Chenggang Wu ◽

Chengjun Li ◽

Shawn S.-C. Li

Keyword(s):

Rich Region ◽

Binding Motifs ◽

Proteomic Screen ◽

Src Homology 3 ◽

Src Homology 3 Domain ◽

Src Homology

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In vitro characterization of major ligands for Src homology 2 domains derived from protein tyrosine kinases, from the adaptor protein SHC and from GTPase-activating protein in Ramos B cells

European Journal of Immunology ◽

10.1002/eji.1830240812 ◽

1994 ◽

Vol 24 (8) ◽

pp. 1799-1807 ◽

Cited By ~ 48

Author(s):

Götz Baumann ◽

Daniel Maier ◽

Felix Freuler ◽

Claude Tschopp ◽

Kristin Baudisch ◽

...

Keyword(s):

B Cells ◽

Tyrosine Kinases ◽

Adaptor Protein ◽

Protein Tyrosine Kinases ◽

Gtpase Activating Protein ◽

Src Homology 2 ◽

In Vitro Characterization ◽

Src Homology

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Molecular Cloning and Characterization of cgt, the Brucella abortus Cyclic β-1,2-Glucan Transporter Gene, and Its Role in Virulence

Infection and Immunity ◽

10.1128/iai.72.4.2263-2271.2004 ◽

2004 ◽

Vol 72 (4) ◽

pp. 2263-2271 ◽

Cited By ~ 31

Author(s):

Mara S. Roset ◽

Andrés E. Ciocchini ◽

Rodolfo A. Ugalde ◽

Nora Iñón de Iannino

Keyword(s):

Brucella Abortus ◽

Sinorhizobium Meliloti ◽

Nodule Development ◽

Nucleotide Sequencing ◽

Nodule Occupancy ◽

Transporter Gene ◽

Binding Motifs ◽

Reading Frame ◽

Intracellular Multiplication

ABSTRACT The animal pathogen Brucella abortus contains a gene cgt, which complemented Sinorhizobium meliloti nodule development (ndvA) and Agrobacterium tumefaciens chromosomal virulence (chvA) mutants. Complemented strains recovered the presence of anionic cyclic β-1,2-glucan, motility, tumor induction in A. tumefaciens, and nodule occupancy in S. meliloti, all traits strictly associated with the presence of cyclic β-1,2-glucan in the periplasm. Nucleotide sequencing revealed that B. abortus cgt contains a 1,797-bp open reading frame coding for a predicted membrane protein of 599 amino acids (65.9 kDa) that is 58.5 and 59.9% identical to S. meliloti NdvA and A. tumefaciens ChvA, respectively. Additionally, B. abortus cgt, like S. meliloti ndvA and A. tumefaciens chvA possesses ATP-binding motifs and the ABC signature domain features of a typical ABC transporter. Characterization of Cgt was carried out by the construction of null mutants in B. abortus 2308 and S19 backgrounds. Both mutants do not transport cyclic β-1,2-glucan to the periplasm, as shown by the absence of anionic cyclic glucan, and they display reduced virulence in mice and defective intracellular multiplication in HeLa cells. These results suggest that cyclic β-1,2-glucan must be transported into the periplasmatic space to exert its action as a virulence factor.

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Identification and characterization of SPRK, a novel src-homology 3 domain-containing proline-rich kinase with serine/threonine kinase activity.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(17)36578-x ◽

1994 ◽

Vol 269 (21) ◽

pp. 15092-15100 ◽

Cited By ~ 4

Author(s):

K.A. Gallo ◽

M.R. Mark ◽

D.T. Scadden ◽

Z. Wang ◽

Q. Gu ◽

...

Keyword(s):

Kinase Activity ◽

Serine Threonine Kinase ◽

Threonine Kinase ◽

Src Homology 3 ◽

Src Homology 3 Domain ◽

Src Homology ◽

Identification And Characterization

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Initial characterization of TREM-like transcript (TLT)–1: a putative inhibitory receptor within the TREM cluster

Blood ◽

10.1182/blood-2002-02-0523 ◽

2002 ◽

Vol 100 (10) ◽

pp. 3822-3824 ◽

Cited By ~ 40

Author(s):

A. Valance Washington ◽

Laura Quigley ◽

Daniel W. McVicar

Keyword(s):

Cell Activation ◽

Bone Marrow Cells ◽

Myeloid Cell ◽

Dendritic Cell Maturation ◽

Chromosome 17 ◽

Inhibitory Receptor ◽

Src Homology 3 ◽

Src Homology 3 Domain ◽

Src Homology

The TREMs (triggering receptors expressed on myeloid cells) represent a family of 5 receptors clustered on murine chromosome 17. TREMs 1 and 2 affect various aspects of myeloid cell activation and development, including responsiveness to lipopolysaccharide and regulation of dendritic cell maturation, yet no inhibitory receptor has been demonstrated within this cluster. Here we characterize TLT-1 (TREM-like transcript-1), a putative inhibitory receptor within the TREM cluster that contains an extracellular V-set Ig domain, a proline-rich region, and an immune receptor tyrosine-based inhibitory motif (ITIM) in its cytoplasmic tail. To our knowledge, TLT-1 is the first ITIM-containing receptor carrying a potential Src homology 3 domain ligand. TLT-1 transcripts are abundant in bone marrow cells, but not in lymphocytes, and phosphorylated TLT-1 associates with SHP-1, suggesting that it is indeed an inhibitory receptor. Based on these characteristics, it is likely that TLT-1 regulates the signaling of the TREM family receptors.

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