Identification of a novel ubiquitin conjugation motif, required for ligand-induced internalization of the growth hormone receptor.
In addition to its role in selective protein degradation, the conjugation of ubiquitin to proteins has also been implicated in the internalization of plasma membrane proteins, including the alpha-factor receptor Ste2p, uracil permease Fur4p, epithelial sodium channel ENaC and the growth hormone receptor (GHR). Binding of GH to its receptor induces receptor dimerization, resulting in the activation of signal transduction pathways and an increase of GHR ubiquitination. Previously, we have shown that the ubiquitin conjugation system mediates GH-induced GHR internalization. Here, we present evidence that a specific domain of the GHR regulates receptor endocytosis via the ubiquitin conjugation system. This ubiquitin-dependent endocytosis (UbE) motif consists of the amino acid sequence DSWVEFIELD and is homologous to sequences in other proteins, several of which are known to be ubiquitinated. In addition, we show that GH internalization by a truncated GHR is independent of the presence of lysine residues in the cytosolic domain of this receptor, while internalization still depends on an intact ubiquitin conjugation system. Thus, GHR internalization requires the recruitment of the ubiquitin conjugation system to the GHR UbE motif rather than the conjugation of ubiquitin to the GHR itself.
Govers
R
R
Department of Cell Biology, Faculty of Medicine and Institute of Biomembranes, Utrecht University, Utrecht, The Netherlands.
ten Broeke
T
T
van Kerkhof
P
P
Schwartz
A L
AL
Strous
G J
GJ
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D016466:CHO Cells; D006224:Cricetinae; D004705:Endocytosis; D013006:Growth Hormone / Q000378:metabolism; D008024:Ligands; D008239:Lysine / Q000737:chemistry; D009154:Mutation; D011817:Rabbits; D011986:Receptors, Somatotropin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014452:Ubiquitins / Q000378:metabolism*
D008024:Ligands; D011986:Receptors, Somatotropin; D011994:Recombinant Proteins; D014452:Ubiquitins; D013006:Growth Hormone; D008239:Lysine