Intracellular localization of homopolymeric amino acid-containing proteins expressed in mammalian cells.
Many human proteins have homopolymeric amino acid (HPAA) tracts, which are involved in protein-protein interactions and also have intrinsic polymerization properties. Polyglutamine or polyalanine expansions cause several neurodegenerative diseases. To examine the properties of HPAAs, we expressed 20 kinds of 30-residue HPAA fused to the C terminus of yellow fluorescent protein in mammalian cells. Specific localization was observed depending on the HPAA. Polyarginine and polylysine aggregated in the nucleus. Polyalanine, polyhistidine, polyisoleucine, polyleucine, polymethionine, polyphenylalanine, polythreonine, polytryptophan, and polyvaline localized in the cytoplasm, and some of these HPAAs formed aggregate(s). Hydrophobic HPAAs such as polyisoleucine, polyleucine, polyphenylalanine, and polyvaline were found as one major aggregate or cumulus in the perinuclear region. Western blot analysis indicated that hydrophobic HPAA tracts appear to oligomerize and form high molecular weight complexes. These results indicate that hydrophobicity itself may trigger the oligomerization and aggregation of proteins when overexpressed in cells. Our experiments provide novel insights into the nature of the HPAAs that are often seen in human and other organisms.
Oma
Yoko
Y
Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Tokyo 153-8902, Japan.
Kino
Yoshihiro
Y
Sasagawa
Noboru
N
Ishiura
Shoichi
S
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D019556:COS Cells; D002522:Chlorocebus aethiops; D017930:Genes, Reporter; D015894:Genome, Human; D006801:Humans; D042541:Intracellular Space; D008164:Luminescent Proteins / Q000235:genetics; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000378:metabolism; D014162:Transfection
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D010455:Peptides; D011506:Proteins; C054562:yellow fluorescent protein, Bacteria