Highly preferential nucleation of histone H1 assembly on scaffold-associated regions

Izaurralde, Elisa; Käs, Emmanuel; Laemmli, Ulrich Karl

doi:10.1016/0022-2836(89)90133-2

Cited by 165 publications

(106 citation statements)

References 41 publications

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“…The membranes were blocked for 2 h in Blotto (20 mM HEPES, pH 7.5, 25 mM NaCl, 5 mM MgCl 2 , 1 mM dithiothreitol, 0.05% Nonidet P-40, 5% skimmed milk powder) at room temperature followed by an incubation in Hyb 75 (20 mM HEPES, pH 7.5, 75 mM KCl, 2.5 mM MgCl 2 , 0.1 mM EDTA, 1 mM dithiothreitol, 0.05% Nonidet P-40, 1% skimmed milk powder) for 15 min at room temperature. 35 S-Labeled SAF-B and lamin C were synthesized using the TNT TM T7 Coupled Reticulocyte Lysate System (Promega). The probes were diluted in 3 ml of Hyb 75, and the membranes were incubated for 5 h at 4°C.…”

Section: Methodsmentioning

confidence: 99%

“…In Vitro Binding Assays, Co-immunoprecipitation, and Immunoblotting- 35 S-Labeled, c-Myc-tagged fragments of ZO-2 (PDZ-1 and PDZ-2) and SAF-B (C-term.SAF-B) were synthesized using the TNT TM T7-coupled reticulocyte lysate system (Promega). Equal amounts of protein were combined and incubated at 30°C for 1.5 h followed by the addition of 500 l of co-immunoprecipitation buffer (0.2% Triton X-100, 20 mM Tris⅐Cl, pH 7.5, 150 mM NaCl, 1 mM CaCl 2, 5 mM Na 3 VO 4 , 10 mM NaF, 5 g/ml aprotinin, 5 g/ml leupeptin, 2 g/ml pepstatin A, 1 mM Pefabloc SC, 10% glycerol) and 2 g of mouse monoclonal anti-c-Myc.…”

Section: Methodsmentioning

confidence: 99%

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The Tight Junction Protein ZO-2 Localizes to the Nucleus and Interacts with the Heterogeneous Nuclear Ribonucleoprotein Scaffold Attachment Factor-B

Traweger¹,

Fuchs²,

Krizbai³

et al. 2003

Journal of Biological Chemistry

141

120

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Zonula occludens proteins (ZOPs), currently comprising ZO-1, ZO-2, and ZO-3, belong to the family of membrane-associated guanylate kinase homologue (MAGUK) proteins that are involved in the organization of epithelial and endothelial intercellular junctions. ZOPs bind to the cytoplasmic C termini of junctional transmembrane proteins linking them to the actin cytoskeleton. They are characterized by several conserved modules, including three PDZ domains, one SH3 domain, and a guanylate kinase-like domain, elements indicating that ZOPs may serve multiple purposes. Interestingly, ZOPs contain some unique motifs not shared by other MAGUK family members, including nuclear localization and nuclear export signals and a leucine zipper-like sequence. Their potential involvement in cell growth and proliferation has been suggested earlier based on the observation that the N-terminal half of ZOPs displays significant similarity to the product of the Drosophila tumor suppressor gene lethal(1)discslarge (dlg). The nuclear targeting of ZOPs in subconfluent epithelial cell cultures is well documented, although the action of the junctional MAGUKs in the nucleus has remained elusive. Here we show for the first time that nuclear ZO-2 directly interacts with the DNA-binding protein scaffold attachment factor-B (SAF-B). Our results from two-hybrid assays and in vivo co-immunoprecipitation studies provide evidence to suggest that ZO-2 associates with the C-terminal portion of SAF-B via its PDZ-1 domain. We further demonstrate that enhanced green fluorescent protein (EGFP)-and DsRed-tagged ZO-2 and SAF-B fusion proteins partially co-localize in nuclei of transfected epithelial cells. As shown by laser confocal microscopy and epifluorescent analysis, nuclear ZO-2 is present in epithelial and endothelial cells, particularly in response to environmental stress conditions. Interestingly, no association of SAF-B with ZO-1 was found, which supports the notion that junctional MAGUKs serve nonredundant functions. Tight intercellular contacts (tight junctions (TJs))1 are responsible for the "barrier" function of epithelia and endothelia restricting the paracellular transport of solutes and molecules across epithelial and endothelial cell sheets (1-5). TJs consist of several transmembrane components that interact with a network of "peripheral" cytoplasmic proteins. So far, three types of TJ-related transmembrane proteins: occludin, claudins, and junctional adhesion molecule, have been described, all of them associating with at least one of the Zonula occludens proteins (ZOPs) (for review see Refs. 6 -8). ZOPs, in turn, establish a link between the junction site and the cytoskeleton by interacting directly with actin filaments (9 -11). The observation that ZOPs not only associate with each other but also with components of adherens junctions and gap junctions in cells lacking TJs (12-14) suggests a universal role for ZOPs at the cytoplasmic surface of the junctional plaque.ZOPs are structurally similar to the membrane-associated guanylate kinase h...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

The Tight Junction Protein ZO-2 Localizes to the Nucleus and Interacts with the Heterogeneous Nuclear Ribonucleoprotein Scaffold Attachment Factor-B

Traweger¹,

Fuchs²,

Krizbai³

et al. 2003

Journal of Biological Chemistry

141

120

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“…The 5h end is ' capped ' by the addition of a methylated guanidine residue after approx. 30 nucleotides have been synthesised [19,20]. The 5h cap is an important signal for the selection of the start AUG by the ribosome complex during initiation of translation, and also appears to protect the RNA molecule from 5h-3h degradation [21,22].…”

Section: Post-transcriptional Regulationmentioning

confidence: 99%

“…During eukaryotic interphase, chromosomes are organized into looped domains which are anchored to the nuclear matrix through DNA regions called matrix-attachment regions (MARs) [19,86]. MAR-binding proteins generally recognize and bind to the minor groove or the intrinsic bend of AT-rich DNA [87], and subsequently may function as important transcriptional control proteins during interphase.…”

Section: Classical Silencers (Silencer Elements)mentioning

confidence: 99%

Transcriptional control and the role of silencers in transcriptional regulation in eukaryotes

Ogbourne

Antalis

1998

Biochemical Journal

222

171

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Mechanisms controlling transcription and its regulation are fundamental to our understanding of molecular biology and, ultimately, cellular biology. Our knowledge of transcription initiation and integral factors such as RNA polymerase is considerable, and more recently our understanding of the involvement of enhancers and complexes such as holoenzyme and mediator has increased dramatically. However, an understanding of transcriptional repression is also essential for a complete understanding of promoter structure and the regulation of gene expression. Transcriptional repression in eukaryotes is achieved through ‘silencers ’, of which there are two types, namely ‘silencer elements ’ and ‘negative regulatory elements ’ (NREs). Silencer elements are classical, position-independent elements that direct an active repression mechanism, and NREs are position-dependent elements that direct a passive repression mechanism. In addition, ‘repressors ’ are DNA-binding trasncription factors that interact directly with silencers. A review of the recent literature reveals that it is the silencer itself and its context within a given promoter, rather than the interacting repressor, that determines the mechanism of repression. Silencers form an intrinsic part of many eukaryotic promoters and, consequently, knowledge of their interactive role with enchancers and other transcriptional elements is essential for our understanding of gene regulation in eukaryotes.

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“…Transient transfections such as these do not represent the physiological scenario where histone proteins and architectural elements of tertiary structure, such as scaffold attachment regions (SARs) are involved. An indicator of an SAR is the presence of AT-rich elements, 35,36 and such a region is located 5Ј of −1025hLuc. It is possible that complex secondary structure resulting from internal homology is formed at this region, preventing TF ingress, and thus transcriptional activation.…”

Section: Discussionmentioning

confidence: 99%

Design of a muscle cell-specific expression vector utilising human vascular smooth muscle α-actin regulatory elements

Chen

et al. 1999

Gene Ther

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Highly preferential nucleation of histone H1 assembly on scaffold-associated regions

Cited by 165 publications

References 41 publications

The Tight Junction Protein ZO-2 Localizes to the Nucleus and Interacts with the Heterogeneous Nuclear Ribonucleoprotein Scaffold Attachment Factor-B

The Tight Junction Protein ZO-2 Localizes to the Nucleus and Interacts with the Heterogeneous Nuclear Ribonucleoprotein Scaffold Attachment Factor-B

Transcriptional control and the role of silencers in transcriptional regulation in eukaryotes

Design of a muscle cell-specific expression vector utilising human vascular smooth muscle α-actin regulatory elements

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