Fischbach Lab, Information about UAS-HB3 line

Targeted expression of IrreC-rst in central complex structures by use of theGAL4/UAS-System

Reiter, C., Nie, Z. & Fischbach, K.-F.
Institut für Biologie III, Schänzlestr.1, D-79104 Freiburg

IrreC-rst (see below), a transmembrane protein of the immunoglobulin superfamily with 5immunoglobulin-like domains, is required for normal axonal projections in both optic chiasms(Boschert et al., 1990; Ramos et al., 1993). Normally it is not expressed in developing centralcomplex structures. When misexpressed in these structures (by the use of the Gal4/UAS-system;Brand and Perrimon, 1993), it does not seem to affect their development. We cloned the irreC-rstcDNA HB3 into the pUAST vector, behind UAS binding sites for the yeast transcription factor Gal4and established five independent UAS-HB3 transformant lines, that were subsequently crossed toGal4-lines.

The IrreC-rst protein in F1 hybrids was detected by mab24A5.1 which is directed against the extracellular domain of IrreC-rst (Schneider et al., 1995). We viewed pupal brain whole mounts in the confocal microscope and scanned in 1024 x 1024 mode.

Sequence comparison with DM-GRASP (chicken) and NEUROLIN (goldfish)

The similarity of the IrreC-rst protein to its vertebrate relatives is best in the extracellular domain (with the exception of the N-terminal region). TM denotes the transmembrane domains. Assuming a more primitive structure of the IrreC-rst protein, the intracellular domains of the vertebrate molecules might have evolved by several deletions.

DM-GRASP sequence (chicken) from:

Burns, F.R., Von Kannen, S., Guy, L., Raper, J.A., Kamholz, J., and Chang, S. (1991). DM-GRASP, a novel immunoglobulin superfamily axonal surface protein that supports neurite extension. Neuron 7, 209-220

Neurolin sequence (goldfish) from:

Laessing, U., Giordano, S., Stecher, B., Lottspeich, F., and Stuermer, C.A.O. (1994). Molecular characterization of fish neurolin: a growth-associated cell surface protein and member of the immunoglobulin superfamily in the fish retinotectal system with similarities to chick protein DM-GRASP/SC-1/BEN. Differentiation 56, 21-29

IrreC-rst sequence (Drosophila) from:

Ramos, R.G.P., Igloi, G.L., Lichte, B., Baumann, U., Maier, D., Schneider, T., Brandstätter, J.H., Fröhlich, A., and Fischbach, K.-F. (1993). The irregular chiasma C - roughest locus of Drosophila, which affects axonal projections and programmed cell death, encodes a novel immunoglobulin-like protein. Genes and Development 7, 2533-2547

References

Brand, A.H., Perrimon, N. (1993). Development 118:401-415

Ramos, R. C., Igloi, G. L., Lichte, B., Baumann, U., Maier, D., Schneider, T., Brandstätter, J. H., Fröhlich, A., and Fischbach, K. F. (1993). Genes Dev. 7, 2533-2547.

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