Visualization of the interaction between histone H3 and lamin B1 using Navinci’s in situ proximity ligation technology strengthens our understanding of the integrity of chromatin architecture and dynamics in human cells, particularly in the context of cancer progression and cell fate.
Exploring nucleus and chromatin dynamics: protein-protein interaction histone H3 and lamin B1
Visualization of the interaction between histone H3 and lamin B1 using Navinci’s in situ proximity ligation technology strengthens our understanding of the integrity of chromatin architecture and dynamics in human cells, particularly in the context of cancer progression and cell fate.
Histone H3 and lamin B1 interaction in MCF7 cells, detected using NaveniFlex™ Cell Red
The nuclear architecture
The nucleus is a membrane-bound organelle which contains DNA and DNA-associated proteins (chromatin). Chromatin is an order of organization of DNA and its associated proteins (histones) into compact structures. DNA is wrapped around octamers composed of two copies each of histones H2A, H2B, H3 and H4 (10), thus forming nucleosomes – the fundamental chromatin subunits. Nucleosomes are further compacted into higher-order structures with the help of histone H1, which in its turn is anchored to lamins (6,7,8,9).
The nuclear envelope is composed of an outer and an inner nuclear membrane (INM), nuclear pore complexes (NPCs) and the nuclear lamina – a proteinaceous meshwork underlying the INM and connected to NPCs (1). This internal nucleoskeleton forms a scaffold involved in chromatin organization and ensures the correct spatial and temporal progression of nuclear processes such as DNA replication and transcription (2, 3, 4). Lamins are major structural components of the scaffold, lining the nuclear periphery where they contribute to the shape and rigidity and of the nucleoskeleton and connect it to the cytoskeleton (5). Lamin B1 has structural domains that directly bind to DNA or histones and indirectly interact with chromatin through LEM proteins [LAP2 (lamina-associated polypeptide 2)-emerin-MAN1] (9). Thus, lamin B1 can provide anchors for chromatin to regulate its position, higher-order structure and dynamics.
Histone H3 and lamin B1 interaction in MCF7 cells, detected using NaveniFlex™ Cell Red
The nuclear architecture
The nucleus is a membrane-bound organelle which contains DNA and DNA-associated proteins (chromatin). Chromatin is an order of organization of DNA and its associated proteins (histones) into compact structures. DNA is wrapped around octamers composed of two copies each of histones H2A, H2B, H3 and H4 (10), thus forming nucleosomes – the fundamental chromatin subunits. Nucleosomes are further compacted into higher-order structures with the help of histone H1, which in its turn is anchored to lamins (6,7,8,9).
The nuclear envelope is composed of an outer and an inner nuclear membrane (INM), nuclear pore complexes (NPCs) and the nuclear lamina – a proteinaceous meshwork underlying the INM and connected to NPCs (1). This internal nucleoskeleton forms a scaffold involved in chromatin organization and ensures the correct spatial and temporal progression of nuclear processes such as DNA replication and transcription (2, 3, 4). Lamins are major structural components of the scaffold, lining the nuclear periphery where they contribute to the shape and rigidity and of the nucleoskeleton and connect it to the cytoskeleton (5). Lamin B1 has structural domains that directly bind to DNA or histones and indirectly interact with chromatin through LEM proteins [LAP2 (lamina-associated polypeptide 2)-emerin-MAN1] (9). Thus, lamin B1 can provide anchors for chromatin to regulate its position, higher-order structure and dynamics.
Figure 1. Model of Navinci’s in situ proximity ligation assay for interaction with histone H3 and lamin B1. Only if the Navenibodies are in close proximity will they generate a rolling circle amplification reaction, leading to a strong and distinct signal
Visualizing histone H3 and lamin B1 interaction
Nuclear morphology plays a vital role in cell fate during cancer progression. Loss of integrity is accompanied by an increased aggressiveness of cancer cells (11). We have demonstrated the use of the classic Naveni® in situ proximity ligation assay for the visualization of the interaction between histone H3 and lamin B1 in both cell lines and FFPE tissues (Figure 1). Furthermore, with Naveni TriFlex Cell we simultaneously demonstrated both the individual expression of histone H3 and lamin B1 and their interaction in different stages of the cell cycle. The lamin B1 and histone H3 interaction is a useful tool for studying the integrity of chromatin architecture and dynamics in human cells.
Visualizing histone H3 and lamin B1 interaction
Nuclear morphology plays a vital role in cell fate during cancer progression. Loss of integrity is accompanied by an increased aggressiveness of cancer cells (11). We have demonstrated the use of the classic Naveni® in situ proximity ligation assay for the visualization of the interaction between histone H3 and lamin B1 in both cell lines and FFPE tissues (Figure 1). Furthermore, with Naveni TriFlex Cell we simultaneously demonstrated both the individual expression of histone H3 and lamin B1 and their interaction in different stages of the cell cycle. The lamin B1 and histone H3 interaction is a useful tool for studying the integrity of chromatin architecture and dynamics in human cells.
Figure 1. Model of Navinci’s in situ proximity ligation assay for interaction with histone H3 and lamin B1. Only if the Navenibodies are in close proximity will they generate a rolling circle amplification reaction, leading to a strong and distinct signal
Application example – Unveiling cellular drama
Witness the cellular saga through Naveni TriFlex Cell, revealing the dynamic interplay of histone H3 and lamin B1 in key cellular events.
Application example – Unveiling cellular drama
Witness the cellular saga through Naveni TriFlex Cell, revealing the dynamic interplay of histone H3 and lamin B1 in key cellular events.
Birth: Mitotic cells exhibit diffuse lamin B1 signals (yellow) during nuclear envelope breakdown, while histone H3 (magenta) remains near the metaphase plate. Histone H3 and lamin B1 complexes (cyan) disassemble.
Life: In non-dividing cells, free lamin B1 localizes in the nuclear envelope, and multiple histone H3 and lamin B1 interactions occur in the nucleoplasm alongside chromatin-bound histone H3.
Death: Early oncosis sees a swollen nucleus with less densely packed complexes in the nucleoplasm, featuring unbound histone H3 and lamin B1.
Birth: Mitotic cells exhibit diffuse lamin B1 signals (yellow) during nuclear envelope breakdown, while histone H3 (magenta) remains near the metaphase plate. Histone H3 and lamin B1 complexes (cyan) disassemble.
Life: In non-dividing cells, free lamin B1 localizes in the nuclear envelope, and multiple histone H3 and lamin B1 interactions occur in the nucleoplasm alongside chromatin-bound histone H3.
Death: Early oncosis sees a swollen nucleus with less densely packed complexes in the nucleoplasm, featuring unbound histone H3 and lamin B1.
How to detect histone H3 and lamin B1
Use our products Naveni TriFlex Cell or NaveniFlex Cell. These products were used in the examples above. For further information about histone/lamin monoclonal antibodies and protocols, contact us using the form below.
How to detect histone H3 and lamin B1
Use our products Naveni TriFlex Cell or NaveniFlex Cell. These products were used in the examples above. For further information about histone/lamin monoclonal antibodies and protocols, contact us using the form below.
Naveni TriFlex Cell
The kit enables fluorescent staining of protein-protein interactions, concurrently visualizing the two proteins in their free state.
NaveniFlex Cell
The assay is designed to be used with a mouse and rabbit primary antibody pair. Detection with TEX615 fluorophore is included.
Naveni TriFlex Cell
The assay is designed to be used with a mouse and a rabbit primary pair. Chromogenic readout with HRP substrate.
NaveniFlex Cell
The assay is designed to be used with a mouse and rabbit primary antibody pair. Detection with TEX615 fluorophore is included.
References
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- Bridger, J. M., Foeger, N., Kill, I. R., & Herrmann, H. (2007). The nuclear lamina. Both a structural framework and a platform for genome organization. The FEBS journal, 274(6), 1354–1361. DOI: 10.1111/j.1742-4658.2007.05694.x
- Dechat, T., Pfleghaar, K., Sengupta, K., Shimi, T., Shumaker, D. K., Solimando, L., & Goldman, R. D. (2008). Nuclear lamins: major factors in the structural organization and function of the nucleus and chromatin. Genes & development, 22(7), 832–853. DOI: 10.1101/gad.1652708
- Dorner, D., Gotzmann, J., & Foisner, R. (2007). Nucleoplasmic lamins and their interaction partners, LAP2alpha, Rb, and BAF, in transcriptional regulation. The FEBS journal, 274(6), 1362–1373. DOI: 10.1111/j.1742-4658.2007.05695.x
- Crisp, M., & Burke, B. (2008). The nuclear envelope as an integrator of nuclear and cytoplasmic architecture. FEBS Letters, 582(14), 2023–2032. DOI: 10.1016/j.febslet.2008.05.001
- Shumaker, D. K., Kuczmarski, E. R., & Goldman, R. D. (2003). The nucleoskeleton: lamins and actin are major players in essential nuclear functions. Current opinion in cell biology, 15(3), 358–366. DOI: 10.1016/s0955-0674(03)00050-4
- Chang, L., Li, M., Shao, S., Li, C., Ai, S., Xue, B., Hou, Y., Zhang, Y., Li, R., Fan, X., He, A., Li, C., & Sun, Y. (2022). Nuclear peripheral chromatin-lamin B1 interaction is required for global integrity of chromatin architecture and dynamics in human cells. Protein & Cell, 13(4), 258–280. DOI: 10.1007/s13238-020-00794-8
- Robinson, P. J., & Rhodes, D. (2006). Structure of the ’30 nm’ chromatin fibre: a key role for the linker histone. Current opinion in structural biology, 16(3), 336–343. DOI: 10.1016/j.sbi.2006.05.007
- Barton, L. J., Soshnev, A. A., & Geyer, P. K. (2015). Networking in the nucleus: a spotlight on LEM-domain proteins. Current opinion in cell biology, 34, 1–8. DOI: 10.1016/j.ceb.2015.03.005
- Mariño-Ramírez, L., Kann, M. G., Shoemaker, B. A., & Landsman, D. (2005). Histone structure and nucleosome stability. Expert review of proteomics, 2(5), 719–729. DOI: 10.1586/14789450.2.5.719
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Fischer, E. G. (2020). Nuclear Morphology and the Biology of Cancer Cells. Acta Cytologica, 64(6), 511–519. DOI: 10.1159/000508780