Aggregation independent of N-cadherin and neural cell adhesion molecule on quail myoblasts transformed with temperature-sensitive Rous sarcoma virus.

Abstract

Quail myoblasts transformed with the temperature-sensitive mutant of Rous sarcoma virus (QM-RSV cells) differentiate temperature-sensitively. At 41 degrees C, the cells begin to fuse after about 15-18 h and form multinucleated myotubes, whereas, at 35.5 degrees C, the cells proliferate. Tyrosine-phosphorylation relates to this temperature-sensitive differentiation. In the course of the investigation of QM-RSV cells, when QM-RSV cells were dissociated with EDTA and shaken in DMEM, the aggregation activity was detected. This activity was expressed on the cells cultured at 41 degrees C, but not at 35.5 degrees C. For detailed characterization of the aggregation, cells from which cadherin and/or neural cell adhesion molecule (NCAM) were removed by trypsin treatment were used. It was then observed that temperature-sensitive and calcium-dependent aggregation activity was expressed on the cells treated with trypsin and EDTA (TE-cells), although the TE-cells did not retain either aggregation molecule. The aggregation activity began to be expressed at 2-4 h after temperature shift and increased with the differentiation. The expression of the activity related to the tyrosine-phosphorylation of some protein. The aggregation of TE-cells was completely inhibited by D(+)-mannose, D(+)-glucose, and N-acetyl-D-glucosamine, but D(+)-galactose did not affect the aggregation. Thus, the present results suggest that the aggregation of mannose specific C-type animal lectin recognized on TE-cells relates to the early stage of the differentiation of QM-RSV cells.