Identification and characterization of two novel SH2 domain-containing proteins from a yeast two hybrid screen with the ABL tyrosine kinase

Abstract

To further our understanding of the molecular mechanism of Bcr ± Abl mediated transformation, a yeast two hybrid screen was used to identify proteins binding to the Abl tyrosine kinase. Two partial cDNAs encoding novel SH2 domain-containing proteins were cloned and designated Shd and She. Both have homology to Shb, a previously reported SH2 domain-containing protein.Northern blot analysis showed that She is expressed in heart, lung, brain, and skeletal muscle, while expression of Shd is restricted to the brain. The deduced amino acid sequence of the full length mouse Shd cDNA contains an amino-terminal proline-rich region, and a carboxyterminal SH2 domain. A bacterially expressed Shd domain bound multiple tyrosine-phosphorylated proteins with relative molecular weights of 200, 170, 130, 100, 90, 78, 72 and 32 kDa from K562 cell lysates. Shd contains ®ve YXXP motifs, a substrate sequence preferred by Abl tyrosine kinases. Shd was tyrosine phosphorylated in COS-7 cells co-transfected with Shd and c-Abl or Bcr ± Abl. These results suggest that Shd may be a physiological substrate of c-Abl and may function as an adapter protein in the central nervous system.

A Novel TATA-Binding Protein-Binding Protein, ABT1, Activates Basal Transcription and Has a Yeast Homolog That Is Essential for Growth

“…pB42AD-mABT1, which contains the mouse ABT1 cDNA, was isolated from the Mouse Embryo MATCHMAKER LexA cDNA Library (Clontech) using the SH2 domain of SHD (48). Both strands of the mABT1 cDNA were sequenced with an ABI 377 DNA sequencer (Perkin-Elmer).…”

“…mABT1 was isolated during the course of yeast two-hybrid screening using the Src homology 2 (SH2) domain of SHD, a previously identified SH2 domain-containing protein (48). The analysis carried out in this study showed that mABT1 associated directly with TBP and activated transcription from an adenovirus major late (AdML) promoter in a cell-free system.…”

The role of the Src Homology-2 domain containing protein B (SHB) in β cells

“…SHB has three paralogues: SHD, SHE and SHF (Oda et al 1997, Lindholm et al 2000. They all share sequence similarity in the C-terminus and at the phosphorylation sites.…”

“…In addition, SHD exhibits limited sequence similarity with SHB in the N-terminus. SHD and SHE were identified as c-Abl interacting partners (Oda et al 1997), whereas SHF was characterized based on sequence homology with SHB (Lindholm et al 2000). Whereas little is known about the functions of SHD and SHE, SHF has been reported to negatively regulate anaplastic lymphoma kinase (ALK) signaling (Takagi et al 2013) and participate in PDGFRA signaling (Lindholm et al 2000).…”

Efeito da vitamina D no perfil transcricional de cultura organotípica de câncer de mama

“…Frequentemente está desregulado no câncer, sendo hiporegulado ou perdido em alguns tumores e superexpresso em outros (Havre et al,2008;Sulda et al, 2006). Wesley et al (1999) SHE, domínio SH2 contendo a proteína E, constitui uma proteína do domínio de SH2, cujo RNAm foi encontrado em coração, pulmão, cérebro e músculo esquelético de tecido murino (Oda et al, 1997). Como o domínio SH2 pode estar presente em diferentes moléculas de modo a exercer diversos efeitos, é difícil predizer se ocorre uma atuação oncogênica em SH2 e, consequentemente em SHE, como no oncogene Src.…”