scholarly journals Mammalian Protein-Protein Interaction Trap (MAPPIT) Analysis of STAT5, CIS, and SOCS2 Interactions with the Growth Hormone Receptor

2007 ◽  
Vol 21 (11) ◽  
pp. 2821-2831 ◽  
Author(s):  
Isabel Uyttendaele ◽  
Irma Lemmens ◽  
Annick Verhee ◽  
Anne-Sophie De Smet ◽  
Joël Vandekerckhove ◽  
...  
Keyword(s):  
Protein Interaction ◽  
Binding Sites ◽  
Growth Hormone Receptor ◽  
Functional Divergence ◽  
Critical Role ◽  
Sh2 Domain ◽  
Cytokine Signaling ◽  
Protein Protein Interaction ◽  
Interaction Trap ◽  
Mammalian Protein

Abstract Binding of GH to its receptor induces rapid phosphorylation of conserved tyrosine motifs that function as recruitment sites for downstream signaling molecules. Using mammalian protein-protein interaction trap (MAPPIT), a mammalian two-hybrid method, we mapped the binding sites in the GH receptor for signal transducer and activator of transcription 5 (STAT5) a and b and for the negative regulators of cytokine signaling cytokine-inducible Src-homology 2 (SH2)-containing protein (CIS) and suppressor of cytokine signaling 2 (SOCS2). Y534, Y566, and Y627 are the major recruitment sites for STAT5. A non-overlapping recruitment pattern is observed for SOCS2 and CIS with positions Y487 and Y595 as major binding sites, ruling out SOCS-mediated inhibition of STAT5 activation by competition for shared binding sites. More detailed analysis revealed that CIS binding to the Y595, but not to the Y487 motif, depends on both its SH2 domain and the C-terminal part of its SOCS box, with a critical role for the CIS Y253 residue. This functional divergence of the two CIS/SOCS2 recruitment sites is also observed upon substitution of the Y+1 residue by leucine, turning the Y487, but not the Y595 motif into a functional STAT5 recruitment site.

scholarly journals Counter-selectable marker for bacterial-based interaction trap systems

BioTechniques ◽  
2006 ◽  
Vol 40 (2) ◽  
pp. 179-184 ◽  
Author(s):  
Xiangdong Meng ◽  
Robin M. Smith ◽  
Astrid V. Giesecke ◽  
J. Keith Joung ◽  
Scot A. Wolfe
Keyword(s):  
Selectable Marker ◽  
Interaction Trap

scholarly journals Interaction Trap/Two‐Hybrid System to Identify Interacting Proteins

2000 ◽  
Vol 8 (1) ◽  
Author(s):  
Erica A. Golemis ◽  
Ilya Serebriiskii ◽  
Russell L. Finley ◽  
Mikhail G. Kolonin ◽  
Jeno Gyuris ◽  
...  
Keyword(s):  
Hybrid System ◽  
Interacting Proteins ◽  
Two Hybrid ◽  
Interaction Trap

scholarly journals Functional Interaction Trap

2003 ◽  
Vol 2 (11) ◽  
pp. 1217-1224 ◽  
Author(s):  
Alok Sharma ◽  
Susumu Antoku ◽  
Kosaku Fujiwara ◽  
Bruce J. Mayer
Keyword(s):  
Functional Interaction ◽  
Interaction Trap

scholarly journals MAPPIT: a versatile tool to study cytokine receptor signalling

2008 ◽  
Vol 36 (6) ◽  
pp. 1448-1451 ◽  
Author(s):  
Irma Lemmens ◽  
Sam Lievens ◽  
Jan Tavernier
Keyword(s):  
Protein Interactions ◽  
Mammalian Cells ◽  
Cytokine Receptor ◽  
Bait Protein ◽  
Protein Protein Interaction ◽  
Receptor Signalling ◽  
Versatile Tool ◽  
Two Hybrid ◽  
Interaction Trap ◽  
Mammalian Protein

MAPPIT (mammalian protein–protein interaction trap) is a cytokine receptor-based two-hybrid method that operates in intact mammalian cells. A bait is fused C-terminally to a STAT (signal transducer and activator of transcription) recruitment-deficient receptor, whereas the prey is linked to functional STAT-binding sites. When bait and prey interact a ligand-dependent complementation of the STAT recruitment deficiency occurs, leading to activation of a STAT-responsive reporter. MAPPIT is very well suited to study protein interactions involving activated cytokine receptors as the technique allows modification of the bait protein in a physiologically optimal environment.

Spire contains actin binding domains and is related to ascidian posterior end mark-5

Development ◽  
1999 ◽  
Vol 126 (23) ◽  
pp. 5267-5274 ◽  
Author(s):  
A. Wellington ◽  
S. Emmons ◽  
B. James ◽  
J. Calley ◽  
M. Grover ◽  
...  
Keyword(s):  
Actin Cytoskeleton ◽  
Posterior Pole ◽  
Actin Binding ◽  
Binding Domains ◽  
Trap System ◽  
Rho Family Gtpases ◽  
Rho Family ◽  
Interaction Trap ◽  
Anterior Posterior

Spire is a maternal effect locus that affects both the dorsal-ventral and anterior-posterior axes of the Drosophila egg and embryo. It is required for localization of determinants within the developing oocyte to the posterior pole and to the dorsal anterior corner. During mid-oogenesis, spire mutants display premature microtubule-dependent cytoplasmic streaming, a phenotype that can be mimicked by pharmacological disruption of the actin cytoskeleton with cytochalasin D. Spire has been cloned by transposon tagging and is related to posterior end mark-5, a gene from sea squirts that encodes a posteriorly localized mRNA. Spire mRNA is not, however, localized to the posterior pole. SPIRE also contains two domains with similarity to the actin monomer-binding WH2 domain, and we demonstrate that SPIRE binds to actin in the interaction trap system and in vitro. In addition, SPIRE interacts with the rho family GTPases RHOA, RAC1 and CDC42 in the interaction trap system. Thus, our evidence supports the model that SPIRE links rho family signaling to the actin cytoskeleton.

Sign in / Sign up

Export Citation Format

Share Document