Oncogenic EWS-Fli1 interacts with hsRPB7, a subunit of human RNA polymerase II.

As a result of the t(11;22)(q24;q12) chromosomal translocation characterizing the Ewing family of tumors (ET), the amino terminal portion of EWS, an RNA binding protein of unknown function, is fused to the DNA-binding domain of the ets transcription factor Fli1. The hybrid EWS-Fli1 protein acts as a strong transcriptional activator and, in contrast to wildtype Fli1, is a potent transforming agent. Similar rearrangements involving EWS or the highly homologous TLS with various transcription factors have been found in several types of human tumors.

A recombination based method to rapidly assess specificity of two-hybrid clones in yeast.

The yeast two-hybrid system is frequently used to identify protein-protein interactions. Confirming the specificity of candidate clones requires separation and isolation of yeast plasmids, propagation in bacteria and testing combinations of DNA-binding and activation domain hybrids in yeast. In order to simplify this procedure, we developed a rapid method based on PCR amplification of library insert DNAs and in vivo cloning into the activation domain hybrid vector.

Immunoaffinity purification of gamma-aminobutyric acidA (GABAA) receptors containing gamma 1-subunits. Evidence for the presence of a single type of gamma-subunit in GABAA receptors.

Two polyclonal antibodies were raised against the gamma 1-subunit of gamma-aminobutyric acidA (GABAA) receptors. One of these antibodies could be used for the immunopurification of GABAA receptors containing gamma 1-subunits. These receptors exhibited [3H]muscimol and [3H]flunitrazepam binding, and their benzodiazepine binding properties were dramatically different from those of receptors precipitated by anti-gamma 2- or anti-gamma 3-antibodies.

gamma-Aminobutyric acidA receptors displaying association of gamma 3-subunits with beta 2/3 and different alpha-subunits exhibit unique pharmacological properties.

Two polyclonal antibodies were raised against the gamma 3-subunit of gamma-aminobutyric acidA (GABAA) receptors. These antibodies were able to precipitate GABAA receptors from brain membrane extracts, and the precipitated receptors exhibited benzodiazepine binding properties that were dramatically different from those of receptors precipitated by anti-gamma 2 antibodies. The anti-gamma 3 antibodies were used for immunopurification of GABAA receptors containing gamma 3-subunits.