A Di-leucine signal in the ubiquitin moiety. Possible involvement in ubiquitination-mediated endocytosis.
Some plasma membrane receptors in yeast are known to be internalized and degraded in lysosomes upon ligand-dependent ubiquitination. However, the role of ubiquitination in endocytosis and lysosomal degradation in higher eukaryotes has been controversial. In order to directly assess this question, we investigated the fate of chimeric molecules in which ubiquitin moiety was fused in-frame to the cytoplasmic region of membrane proteins. The chimeric proteins with the wild-type ubiquitin were endocytosed and delivered to lysosomes efficiently. Mutant ubiquitin with lysine-to-arginine substitution could still mediate endocytosis, suggesting that polyubiquitination is not required for the endocytosis. We next searched for the existence of an endocytosis signal(s) in the ubiquitin moiety, and identified a di-leucine signal, Leu(43)-Ile(44). The Leu(43)-Ile(44) sequence mediated endocytosis and lysosomal sorting in a Leu(43)-dependent manner. These results suggest that the di-leucine signal in ubiquitin can be involved in ubiquitination-mediated endocytosis and lysosomal targeting of membrane proteins.
Nakatsu
F
F
Department of Molecular Genetics, Chiba University Graduate School of Medicine, Chiba 260-8670, Japan.
Sakuma
M
M
Matsuo
Y
Y
Arase
H
H
Yamasaki
S
S
Nakamura
N
N
Saito
T
T
Ohno
H
H
D000595:Amino Acid Sequence; D000818:Animals; D000944:Antigens, Differentiation, B-Lymphocyte / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004705:Endocytosis; D006367:HeLa Cells; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D006801:Humans; D007532:Isoleucine / Q000378:metabolism; D007930:Leucine / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D008969:Molecular Sequence Data; D015375:Receptors, Interleukin-2 / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction; D014452:Ubiquitins / Q000235:genetics / Q000378:metabolism*; D014633:Valine / Q000378:metabolism
D000944:Antigens, Differentiation, B-Lymphocyte; D000949:Histocompatibility Antigens Class II; D015375:Receptors, Interleukin-2; D011993:Recombinant Fusion Proteins; D014452:Ubiquitins; C045084:invariant chain; D007532:Isoleucine; D007930:Leucine; D014633:Valine