ABSTRACT
Transcription of eukaryotic cell is a multistep process tightly controlled by concerted action of macromolecules. Nuclear receptors are ligand-activated sequence-specific transcription factors that bind DNA and activate (or repress) transcription of specific sets of nuclear target genes. Successful activation of transcription by nuclear receptors and most other transcription factors requires "coregulators" of transcription. Coregulators make up a diverse family of proteins that physically interact with and modulate the activity of transcription factors and other components of the gene expression machinery via multiple biochemical mechanisms. The coregulators include coactivators that accomplish reactions required for activation of transcription and corepressors that suppress transcription. This review summarizes our current knowledge of nuclear receptor coactivators with an emphasis on their biochemical mechanisms of action and means of regulation.
Subject(s)
Nuclear Receptor Coactivators/chemistry , Nuclear Receptor Coactivators/metabolism , Animals , Gene Expression Regulation/physiology , Histone Acetyltransferases/metabolism , Histones/metabolism , Humans , Mice , Nuclear Receptor Coactivators/classification , Receptors, Cytoplasmic and Nuclear/chemistry , Receptors, Cytoplasmic and Nuclear/metabolism , Trans-Activators/metabolismABSTRACT
Most serpins are associated with protease inhibition, and their ability to form loop-sheet polymers is linked to conformational disease and the human serpinopathies. Here we describe the structural and functional dissection of how a unique serpin, the non-histone architectural protein, MENT (Myeloid and Erythroid Nuclear Termination stage-specific protein), participates in DNA and chromatin condensation. Our data suggest that MENT contains at least two distinct DNA-binding sites, consistent with its simultaneous binding to the two closely juxtaposed linker DNA segments on a nucleosome. Remarkably, our studies suggest that the reactive centre loop, a region of the MENT molecule essential for chromatin bridging in vivo and in vitro, is able to mediate formation of a loop-sheet oligomer. These data provide mechanistic insight into chromatin compaction by a non-histone architectural protein and suggest how the structural plasticity of serpins has adapted to mediate physiological, rather than pathogenic, loop-sheet linkages.
Subject(s)
Chromatin/metabolism , DNA-Binding Proteins/chemistry , Models, Molecular , Serpins/chemistry , Animals , Binding Sites , Cathepsin L , Cathepsins/chemistry , Crystallography, X-Ray , Cysteine Endopeptidases/chemistry , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Humans , Mutation , Nucleosomes/metabolism , Protein Conformation , Serpins/genetics , Serpins/metabolismABSTRACT
Posttranslational histone modifications and histone variants form a unique epigenetic landscape on mammalian chromosomes where the principal epigenetic heterochromatin markers, trimethylated histone H3(K9) and the histone H2A.Z, are inversely localized in relation to each other. Trimethylated H3(K9) marks pericentromeric constitutive heterochromatin and the male Y chromosome, while H2A.Z is dramatically reduced at these chromosomal locations. Inactivation of a lysosomal and nuclear protease, cathepsin L, causes a global redistribution of epigenetic markers. In cathepsin L knockout cells, the levels of trimethylated H3(K9) decrease dramatically, concomitant with its relocation away from heterochromatin, and H2A.Z becomes enriched at pericentromeric heterochromatin and the Y chromosome. This change is also associated with global relocation of heterochromatin protein HP1 and histone H3 methyltransferase Suv39h1 away from constitutive heterochromatin; however, it does not affect DNA methylation or chromosome segregation, phenotypes commonly associated with impaired histone H3(K9) methylation. Therefore, the key constitutive heterochromatin determinants can dynamically redistribute depending on physiological context but still maintain the essential function(s) of chromosomes. Thus, our data show that cathepsin L stabilizes epigenetic heterochromatin markers on pericentromeric heterochromatin and the Y chromosome through a novel mechanism that does not involve DNA methylation or affect heterochromatin structure and operates on both somatic and sex chromosomes.
Subject(s)
Cathepsins/metabolism , Centromere/genetics , Cysteine Endopeptidases/metabolism , Heterochromatin/metabolism , Histones/metabolism , Thermodynamics , Y Chromosome/metabolism , Animals , Cathepsin L , Cathepsins/antagonists & inhibitors , Cathepsins/deficiency , Cell Nucleus/metabolism , Chromatin/genetics , Chromosome Segregation/genetics , Chromosomes, Mammalian/genetics , Cysteine Endopeptidases/deficiency , DNA Methylation , Epigenesis, Genetic , Fibroblasts/cytology , Gene Expression , Genetic Markers , Heterochromatin/genetics , Humans , Lysine/metabolism , Male , Methyltransferases/genetics , Methyltransferases/metabolism , Mice , Mice, Knockout , NIH 3T3 Cells , Repressor Proteins/genetics , Repressor Proteins/metabolism , Y Chromosome/geneticsABSTRACT
All cells that constitute mature tissues in an eukaryotic organism undergo a multistep process of cell differentiation. At the terminal stage of this process, cells either cease to proliferate forever or rest for a very long period of time. During terminal differentiation, most of the genes that are required for cell 'housekeeping' functions, such as proto-oncogenes and other cell-cycle and cell proliferation genes, become stably repressed. At the same time, nuclear chromatin undergoes dramatic morphological and structural changes at the higher-order levels of chromatin organization. These changes involve both constitutively inactive chromosomal regions (constitutive heterochromatin) and the formerly active genes that become silenced and structurally modified to form facultative heterochromatin. Here we approach terminal cell differentiation as a unique system that allows us to combine biochemical, ultrastructural and molecular genetic techniques to study the relationship between the hierarchy of chromatin higher-order structures in the nucleus and its function(s) in dynamic packing of genetic material in a form that remains amenable to regulation of gene activity and other DNA-dependent cellular processes.
