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Endocrinology Vol. 139, No. 6 2727-2733
Copyright © 1998 by The Endocrine Society

ARTICLES

Stabilization of the Receptor Protein Tyrosine Phosphatase-Like Protein ICA512 in GH4C1 Cells upon Treatment with Estradiol, Insulin, and Epidermal Growth Factor1

Min S. Lee, Ronald Dirkx, Jr., Michele Solimena and Priscilla S. Dannies

Departments of Pharmacology and Internal Medicine, Section of Endocrinology, Yale University School of Medicine, New Haven, Connecticut 06510

Address all correspondence and requests for reprints to: Dr. Priscilla S. Dannies, Department of Pharmacology, 333 Cedar Street, Yale University School of Medicine, New Haven, Connecticut 06510.

Treatment with 1 nM estradiol, 300 nM insulin, and 5 nM epidermal growth factor induces secretory granule accumulation and prolactin storage in GH4C1 rat pituitary tumor cells. The same triple treatment induced more than 6-fold accumulation of both the precursor (100 kDa, pro-ICA512) and the mature forms (60–70 kDa, ICA512 transmembrane fragment) of ICA512, a receptor protein tyrosine phosphatase-like protein that is preferentially localized in secretory granule membranes. Accumulation of ICA512 resembles that of prolactin storage, for the combination of all three, estradiol, insulin, and epidermal growth factor, gave the greatest induction, which was maximal at 4 days. This effect was specific, as the levels of the small GTP-binding protein Rab3, which is also associated with secretory granule membranes, were unaffected by the triple hormone/growth factor treatment. Increased transcription and translation of ICA512 could only partially account for its 6-fold accumulation, as ICA512 messenger RNA and ICA512 synthesis levels were 1.8 ± 0.35- and 1.6 ± 0.17-fold in triple treated GH4C1 cells compared with those in untreated cells, respectively. Pulse-chase procedures showed that pro-ICA512 was more stable in treated cells. These results indicate that the enlargement of the secretory granule compartment results in the stabilization of ICA512 and raise the possibility that trafficking of secretory granules may affect ICA512’s function and vice versa.




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