Localization of proteasomes and proteasomal proteolysis in the mammalian interphase cell nucleus by systematic application of immunocytochemistry.
Proteasomes are ATP-driven, multisubunit proteolytic machines that degrade endogenous proteins into peptides and play a crucial role in cellular events such as the cell cycle, signal transduction, maintenance of proper protein folding and gene expression. Recent evidence indicates that the ubiquitin-proteasome system is an active component of the cell nucleus. A characteristic feature of the nucleus is its organization into distinct domains that have a unique composition of macromolecules and dynamically form as a response to the requirements of nuclear function. Here, we show by systematic application of different immunocytochemical procedures and comparison with signature proteins of nuclear domains that during interphase endogenous proteasomes are localized diffusely throughout the nucleoplasm, in speckles, in nuclear bodies, and in nucleoplasmic foci. Proteasomes do not occur in the nuclear envelope region or the nucleolus, unless nucleoplasmic invaginations expand into this nuclear body. Confirmedly, proteasomal proteolysis is detected in nucleoplasmic foci, but is absent from the nuclear envelope or nucleolus. The results underpin the idea that the ubiquitin-proteasome system is not only located, but also proteolytically active in distinct nuclear domains and thus may be directly involved in gene expression, and nuclear quality control.
Scharf
Andrea
A
Institut für umweltmedizinische Forschung, Heinrich-Heine-University Düsseldorf, 40225, Düsseldorf, Germany.
Rockel
Thomas Dino
TD
von Mikecz
Anna
A
D000111:Acetylcysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D015415:Biomarkers / Q000378:metabolism*; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D019084:Fluorescent Antibody Technique, Indirect; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry / Q000379:methods*; D007399:Interphase / Q000187:drug effects / Q000502:physiology*; D018613:Microscopy, Confocal; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D016707:Tissue Fixation / Q000379:methods
D015415:Biomarkers; D015853:Cysteine Proteinase Inhibitors; C067713:lactacystin; D046988:Proteasome Endopeptidase Complex; D000111:Acetylcysteine