Distinguishing between retention signals and degrons acting in ERAD.
Endoplasmic reticulum-associated degradation (ERAD) eliminates aberrant proteins from the secretory pathway. Such proteins are retained in the endoplasmic reticulum and targeted for degradation by the ubiquitin-proteasome system. Cis-acting motifs can function in ERAD as retention signals, preventing vesicular export from the endoplasmic reticulum, or as degrons, targeting proteins for degradation. Here, we show that microstp, the C-terminal 20-residue tailpiece of the secretory IgM mus heavy chain, functions both as a portable retention signal and as an ERAD degron. Retention of microstp fusions of secreted versions of thyroid peroxidase and yellow fluorescent protein in the endoplasmic reticulum requires the presence of the penultimate cysteine of microstp. In its role as a portable degron, the microstp targets the retained proteins for ERAD but does not serve as an obligatory ubiquitin-conjugation site. Abolishing microstp glycosylation accelerates the degradation of both microstpCys-fused substrates, yet absence of the N-glycan eliminates the requirement for the penultimate cysteine in the retention and degradation of the unglycosylated yellow fluorescent protein. Hence, the dual role played by the microstpCys motif as a retention signal and as a degron can be attributed to distinct elements within this sequence.
Department of Biochemistry, George S. Wise Faculty of Life Sciences, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel.
Ilana
I
Shapira
Dana
D
Charuvi
Yechiel
Y
Elkabetz
Koret
K
Hirschberg
Shoshana
S
Bar-Nun
D020816:Amino Acid Motifs; D019943:Amino Acid Substitution; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D006031:Glycosylation; D006367:HeLa Cells; D006801:Humans; D007075:Immunoglobulin M / Q000737:chemistry / Q000378:metabolism*; D007148:Immunoglobulin mu-Chains / Q000737:chemistry / Q000378:metabolism*; D007453:Iodide Peroxidase / Q000378:metabolism; D008164:Luminescent Proteins / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D021382:Protein Sorting Signals / Q000502:physiology*; D021381:Protein Transport; D011506:Proteins / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D025801:Ubiquitin / Q000378:metabolism
D007075:Immunoglobulin M; D007148:Immunoglobulin mu-Chains; D008164:Luminescent Proteins; D011134:Polysaccharides; D021382:Protein Sorting Signals; D011506:Proteins; D011993:Recombinant Fusion Proteins; D025801:Ubiquitin; C033027:secretory IgM; D007453:Iodide Peroxidase; D046988:Proteasome Endopeptidase Complex