Proteasomes mediate prolactin-induced receptor down-regulation and fragment generation in breast cancer cells.
Prolactin regulates a variety of physiological processes, including mammary gland growth and differentiation, and recent findings support an important role in breast cancer development and progression. However, little is known about the trafficking of its receptor, a member of the cytokine receptor superfamily. In the present study, we examined the effect of ligand on the endogenous "long" isoform of the prolactin receptor in breast cancer cells. We found that prolactin caused rapid and prolonged down-regulation of this receptor. The prolactin-induced increase in degradation was blocked by inhibitors of both proteasomes and lysosomes. However, the ubiquitin-conjugating system was not required for internalization. Prolactin also resulted in the concomitant appearance of a cell-associated prolactin receptor fragment containing the extracellular domain. This latter process required proteasomal, but not metalloprotease, activity, distinguishing it from ectodomain "shedding" of other membrane receptors, which are secreted as binding proteins. The prolactin receptor fragment was labeled by surface biotinylation and independent of protein synthesis. Together, these data indicated that prolactin binding initiates limited proteasomal cleavage of its receptor, generating a cell-associated fragment containing the extracellular domain. Our findings described a new potential mediator of prolactin action and a novel mechanism whereby proteasomes modulate cellular processes.
Lu
Juu-Chin
JC
Department of Comparative Biosciences, University of Wisconsin-Madison, Wisconsin 53706, USA.
Piazza
Timothy M
TM
Schuler
Linda A
LA
D001943:Breast Neoplasms / Q000473:pathology*; D015536:Down-Regulation; D005260:Female; D006801:Humans; D007536:Isomerism; D008024:Ligands; D008247:Lysosomes; D010446:Peptide Fragments; D011388:Prolactin / Q000378:metabolism / Q000502:physiology*; D046988:Proteasome Endopeptidase Complex / Q000502:physiology*; D011956:Receptors, Cell Surface; D011981:Receptors, Prolactin / Q000502:physiology*; D014407:Tumor Cells, Cultured
D008024:Ligands; D010446:Peptide Fragments; D011956:Receptors, Cell Surface; D011981:Receptors, Prolactin; D011388:Prolactin; D046988:Proteasome Endopeptidase Complex