The huntingtin interacting protein HIP1 is a clathrin and alpha-adaptin-binding protein involved in receptor-mediated endocytosis.
The huntingtin interacting protein (HIP1) is enriched in membrane-containing cell fractions and has been implicated in vesicle trafficking. It is a multidomain protein containing an N-terminal ENTH domain, a central coiled-coil forming region and a C-terminal actin-binding domain. In the present study we have identified three HIP1 associated proteins, clathrin heavy chain and alpha-adaptin A and C. In vitro binding studies revealed that the central coiled-coil domain is required for the interaction of HIP1 with clathrin, whereas DPF-like motifs located upstream to this domain are important for the binding of HIP1 to the C-terminal 'appendage' domain of alpha-adaptin A and C. Expression of full length HIP1 in mammalian cells resulted in a punctate cytoplasmic immunostaining characteristic of clathrin-coated vesicles. In contrast, when a truncated HIP1 protein containing both the DPF-like motifs and the coiled-coil domain was overexpressed, large perinuclear vesicle-like structures containing HIP1, huntingtin, clathrin and endocytosed transferrin were observed, indicating that HIP1 is an endocytic protein, the structural integrity of which is crucial for maintenance of normal vesicle size in vivo.
Waelter
S
S
Max-Planck-Institut für Molekulare Genetik, Ihnestrasse 73, D-14195 Berlin (Dahlem), Germany.
Scherzinger
E
E
Hasenbank
R
R
Nordhoff
E
E
Lurz
R
R
Goehler
H
H
Gauss
C
C
Sathasivam
K
K
Bates
G P
GP
Lehrach
H
H
Wanker
E E
EE
D033965:Adaptor Protein Complex alpha Subunits; D000818:Animals; D019556:COS Cells; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D002460:Cell Line; D002966:Clathrin / Q000737:chemistry* / Q000378:metabolism; D022163:Clathrin-Coated Vesicles / Q000378:metabolism; D004268:DNA-Binding Proteins; D004705:Endocytosis / Q000502:physiology*; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D011233:Precipitin Tests; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D014168:Transferrin
D033965:Adaptor Protein Complex alpha Subunits; D002352:Carrier Proteins; D002966:Clathrin; D004268:DNA-Binding Proteins; C073368:HIP1 protein, human; D008565:Membrane Proteins; D014168:Transferrin