Hsiang‐Cheng Chen

The focal adhesion kinase (FAK) has been implicated in signal transduction pathways initiated by cell adhesion receptor integrins and by neuropeptide growth factors. To gain insight into FAK function, we examined the potential interaction of FAK with intracellular signaling molecules containing the Src homology 2 domains. We report here the stable association of FAK with phosphatidylinositol 3-kinase (PI3-kinase; EC 2.7.1.137) in NIH 3T3 mouse fibroblasts. This interaction was stimulated by cell adhesion concomitant with FAK activation. We also found that recombinant FAK bound to the p85 subunit of PI 3-kinase directly in vitro and that autophosphorylation of recombinant FAK in vitro increased its binding to PI 3-kinase. We detected increased tyrosine phosphorylation of the p85 subunit of PI 3-kinase during cell adhesion and observed direct phosphorylation of p85 by FAK in vitro. Together, these results suggest that PI 3-kinase may be a FAK substrate in vivo and serve as an effector of FAK.

The recently described focal adhesion kinase (FAK) has been implicated in signal transduction pathways initiated by cell adhesion receptor integrins and by neuropeptide growth factors. To examine the mechanisms by which FAK relays signals from the membrane to the cell interior, we carried out a series of experiments to detect potential FAK interactions with proteins containing Src homology 2 (SH2) domains that are important intracellular signaling molecules. Using v-Src-transformed NIH3T3 cells, we showed that FAK was present in the immune-complex precipitated by anti-Src antibody, suggesting potential interaction of FAK with v-Src in vivo. We also showed potentially direct interaction of FAK with v-Src in vivo using the yeast two-hybrid system. Using recombinant FAK expressed in insect cells and bacterial fusion proteins containing Src SH2 domains, we showed direct binding of FAK to the Src SH2 domain but not to the SH3 domain in vitro. A kinase-defective mutant of FAK, which is not autophosphorylated, did not interact with the Src SH2 domain under the same conditions, suggesting the involvement of the FAK autophosphorylation sites. Treatment of FAK with a protein-tyrosine phosphatase decreased its binding to the Src SH2 domain, whereas autophosphorylation in vitro increased its binding. These results confirm the importance of FAK autophosphorylation sites in its interaction with SH2 domain-containing proteins. Taken together, these results suggest that FAK may mediate signal transduction events initiated on the cell surface by kinase activation and autophosphorylation that result in its binding to other key intracellular signaling molecules.

Substances from urinary extracts of normal, nonpregnant subjects and human pituitary gonadotropin preparations were found to react similarly to human chorionic gonadotropin (hCG) in a radioimmunoassay system that is highly specific for hCG and without crossreactivity to human luteinizing hormone (hLH). The antiserum was produced in a rabbit immunized with a bovine albumin conjugate of the unique carboxyl-terminal peptide (residues 123-145) isolated from a tryptic digest of the reduced, S-carboxymethylated hCGbeta subunit. The antibody recognition site on the peptide was found to reside on the last 15 amino acid residues of the carboxyl-terminal peptide, as evidenced by the competitive binding activities against 125I-labeled hCG of a series of peptides chemically synthesized according to the carboxyl-terminal sequence of HCGbeta. In order to elucidate the nature of the crossreacting substance in urinary extracts, a human postmenopausal urinary preparation (Pergonal) and a kaolin-acetone extract of urine from a patient with Klinefelter's syndrome were subjected to gel chromatography on Sephadex G-100. The results indicate that fractions showing immunocrossreactivity with the antiserum to hCGbeta-carboxyl-terminal peptide coeluted with 125I-labeled hCG which was separated distinctly from hLH. The same fractions from this postmenopausal urinary gonadotropin preparation exhibited in vitro biological activity proportional to the immunocrossreactivity of the hCG-specific antiserum. Concentration of postmenopausal women's urine by acetone precipitation retained approximately five times more immunoreactivity per unit volume than kaolin-acetone extraction, when assayed with the antiserum to hCGbeta-carboxyl-terminal peptide.

A herpes-like virus is demonstrated for the first time to be associated with high mortality rates in maricultured abalone Haliotis diversicolor supertexta in Taiwan. Histopathology of moribund abalone indicated that the nerve system was the primary target tissue. The lesions were characterised by tissue necrosis accompanied with infiltration of haemocytes. Electron microscopic examination demonstrated viral particles within the degenerated cerebral ganglion cells. The viruses were hexagonal, approximately 100 nm in diameter and had a single coat. Some viral particles contained a dense nucleoid, while others were empty. The ultrastructure and morphogenesis of the virus particles were consistent with those of the herpesvirus described from the oyster Crassostrea virginica. Experimental infection using supernatant collected from minced visceral organs and muscle of moribund abalone induced 100 % mortality through both intramuscular injection and bath treatments.