title: The yeast two hybrid system in a screen for proteins interacting with axolotl (Ambystoma mexicanum) Msx1 during early limb regeneration.
abstract: Urodele amphibians are exceptional in their ability to regenerate complex body structures such as limbs. Limb regeneration depends on a process called dedifferentiation. Under an inductive wound epidermis terminally differentiated cells transform to pluripotent progenitor cells that coordinately proliferate and eventually redifferentiate to form the new appendage. Recent studies have developed molecular models integrating a set of genes that might have important functions in the control of regenerative cellular plasticity. Among them is Msx1, which induced dedifferentiation in mammalian myotubes in vitro. Herein, we screened for interaction partners of axolotl Msx1 using a yeast two hybrid system. A two hybrid cDNA library of 5-day-old wound epidermis and underlying tissue containing more than 2×10⁶ cDNAs was constructed and used in the screen. 34 resulting cDNA clones were isolated and sequenced. We then compared sequences of the isolated clones to annotated EST contigs of the Salamander EST database (BLASTn) to identify presumptive orthologs. We subsequently searched all no-hit clone sequences against non redundant NCBI sequence databases using BLASTx. It is the first time, that the yeast two hybrid system was adapted to the axolotl animal model and successfully used in a screen for proteins interacting with Msx1 in the context of amphibian limb regeneration.
authors.affiliation: Departmen tof Plastic, Hand and Reconstructive Surgery, Medical School of Hannover, Podbielskistrasse 380, D-30659 Hannover, Germany.
authors.forename: Mehtap
authors.initials: M
authors.lastname: Abuqarn
authors.forename: Christina
authors.initials: C
authors.lastname: Allmeling
authors.forename: Inga
authors.initials: I
authors.lastname: Amshoff
authors.forename: Bjoern
authors.initials: B
authors.lastname: Menger
authors.forename: Inas
authors.initials: I
authors.lastname: Nasser
authors.forename: Peter M
authors.initials: PM
authors.lastname: Vogt
authors.forename: Kerstin
authors.initials: K
authors.lastname: Reimers
mesh_terms: D000558:Ambystoma mexicanum; D000818:Animals; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D005121:Extremities / Q000502:physiology*; D015723:Gene Library; D051957:MSX1 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D012038:Regeneration; D020798:Two-Hybrid System Techniques
chemical_list: D002352:Carrier Proteins; D051957:MSX1 Transcription Factor