TY - JOUR
T1 - Adrm1, a putative cell adhesion regulating protein, is a novel proteasome-associated factor.
AU - Jørgensen, Jakob Ploug
AU - Lauridsen, Anne-Marie
AU - Kristensen, Poul
AU - Dissing, Karen
AU - Johnsen, Anders H
AU - Hendil, Klavs B
AU - Hartmann-Petersen, Rasmus
N1 - Keywords: Amino Acid Sequence; Animals; Cell Adhesion; Cytoplasm; Electrophoresis, Gel, Two-Dimensional; Glycosylation; Hela Cells; Humans; Intracellular Membranes; Membrane Glycoproteins; Mice; Molecular Sequence Data; Organ Specificity; Proteasome Endopeptidase Complex; Protein Binding; Sequence Homology, Amino Acid
PY - 2006
Y1 - 2006
N2 - We have identified Adrm1 as a novel component of the regulatory ATPase complex of the 26 S proteasome: Adrm1 was precipitated with an antibody to proteasomes and vice versa. Adrm1 co-migrated with proteasomes on gel-filtration chromatography and non-denaturing polyacrylamide gel electrophoresis. Adrm1 has been described as an interferon-gamma-inducible, heavily glycosylated membrane protein of 110 kDa. However, we found Adrm1 in mouse tissues only as a 42 kDa peptide, corresponding to the mass of the non-glycosylated peptide chain, and it could not be induced in HeLa cells with interferon. Adrm1 was present almost exclusively in soluble 26 S proteasomes, albeit a small fraction was membrane-associated, like proteasomes. Adrm1 was found in cells in amounts equimolar with S6a, a 26 S proteasome subunit. HeLa cells contain no pool of free Adrm1 but recombinant Adrm1 could bind to pre-existing 26 S proteasomes in cell extracts. Adrm1 may be distantly related to the yeast proteasome subunit Rpn13, mutants of which are reported to display no obvious phenotype. Accordingly, knock-down of Adrm1 in HeLa cells had no effect on the amount of proteasomes, or on degradation of bulk cell protein, or accumulation of polyubiquitinylated proteins. This indicates that Adrm1 has a specialised role in proteasome function.
AB - We have identified Adrm1 as a novel component of the regulatory ATPase complex of the 26 S proteasome: Adrm1 was precipitated with an antibody to proteasomes and vice versa. Adrm1 co-migrated with proteasomes on gel-filtration chromatography and non-denaturing polyacrylamide gel electrophoresis. Adrm1 has been described as an interferon-gamma-inducible, heavily glycosylated membrane protein of 110 kDa. However, we found Adrm1 in mouse tissues only as a 42 kDa peptide, corresponding to the mass of the non-glycosylated peptide chain, and it could not be induced in HeLa cells with interferon. Adrm1 was present almost exclusively in soluble 26 S proteasomes, albeit a small fraction was membrane-associated, like proteasomes. Adrm1 was found in cells in amounts equimolar with S6a, a 26 S proteasome subunit. HeLa cells contain no pool of free Adrm1 but recombinant Adrm1 could bind to pre-existing 26 S proteasomes in cell extracts. Adrm1 may be distantly related to the yeast proteasome subunit Rpn13, mutants of which are reported to display no obvious phenotype. Accordingly, knock-down of Adrm1 in HeLa cells had no effect on the amount of proteasomes, or on degradation of bulk cell protein, or accumulation of polyubiquitinylated proteins. This indicates that Adrm1 has a specialised role in proteasome function.
U2 - 10.1016/j.jmb.2006.06.011
DO - 10.1016/j.jmb.2006.06.011
M3 - Journal article
C2 - 16815440
SN - 0022-2836
VL - 360
SP - 1043
EP - 1052
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 5
ER -