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Nuclear Arc Interacts with the Histone Acetyltransferase Tip60 to Modify H4K12 Acetylation(1,2,3).

Wee CL, Teo S, Oey NE, Wright GD, VanDongen HM, VanDongen AM - eNeuro (2014)

Bottom Line: Although Arc protein is found at synapses, it also localizes to the neuronal nucleus, where its function is less understood.Neuronal activity-induced expression of Arc (1) increases endogenous nuclear Tip60 puncta, (2) recruits Tip60 to PML bodies, and (3) increases histone acetylation of Tip60 substrate H4K12, a learning-induced chromatin modification.These mechanisms point to an epigenetic role for Arc in regulating memory consolidation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Program in Neuroscience and Behavioral Disorders, Duke-NUS Graduate Medical School , Singapore 169857.

ABSTRACT
Arc is an immediate-early gene whose genetic ablation selectively abrogates long-term memory, indicating a critical role in memory consolidation. Although Arc protein is found at synapses, it also localizes to the neuronal nucleus, where its function is less understood. Nuclear Arc forms a complex with the β-spectrin isoform βSpIVΣ5 and associates with PML bodies, sites of epigenetic regulation of gene expression. We report here a novel interaction between Arc and Tip60, a histone-acetyltransferase and subunit of a chromatin-remodelling complex, using biochemistry and super-resolution microscopy in primary rat hippocampal neurons. Arc and βSpIVΣ5 are recruited to nuclear Tip60 speckles, and the three proteins form a tight complex that localizes to nuclear perichromatin regions, sites of transcriptional activity. Neuronal activity-induced expression of Arc (1) increases endogenous nuclear Tip60 puncta, (2) recruits Tip60 to PML bodies, and (3) increases histone acetylation of Tip60 substrate H4K12, a learning-induced chromatin modification. These mechanisms point to an epigenetic role for Arc in regulating memory consolidation.

No MeSH data available.


Segmentation of nuclear substructures. A region of interest containing Arc-YFP puncta in a single neuronal nucleus is segmented to outline the Arc structures using Elements AR. The area of each structure is calculated and the mean and SEM are reported for each nucleus, together with the number of puncta (N).
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f1: Segmentation of nuclear substructures. A region of interest containing Arc-YFP puncta in a single neuronal nucleus is segmented to outline the Arc structures using Elements AR. The area of each structure is calculated and the mean and SEM are reported for each nucleus, together with the number of puncta (N).

Mentions: Fluorescence images were obtained using a motorized inverted wide-field epifluorescence microscope (Nikon Eclipse Ti-E), using 40× and 60× Plan-Apo oil objectives, with numerical apertures of 1.35 and 1.49, respectively. Motorized excitation and emission filter wheels (Ludl Electronics) fitted with a DAPI/CFP/YFP/DsRed quad filter set (#86010, Chroma) were used together with filter cubes for DAPI, CFP, YFP and TxRed, and Cy5 (Chroma) to select specific fluorescence signals. Z-stacks were obtained spanning the entire nucleus and out-of-focus fluorescence was removed using the AutoQuant 3D deconvolution algorithm (Media Cybernetics). Images were digitized using a cooled EM-CCD camera (iXon EM+ 885; Andor). Image acquisition was performed using NIS Elements AR 3.1 software (Nikon). NIS Elements analysis tools were used to outline the nuclei based on their DAPI images and to measure areas of the various nuclear substructures, as illustrated in Figure 1 for Arc-YFP puncta in the nucleus of a 20 DIV hippocampal neuron.


Nuclear Arc Interacts with the Histone Acetyltransferase Tip60 to Modify H4K12 Acetylation(1,2,3).

Wee CL, Teo S, Oey NE, Wright GD, VanDongen HM, VanDongen AM - eNeuro (2014)

Segmentation of nuclear substructures. A region of interest containing Arc-YFP puncta in a single neuronal nucleus is segmented to outline the Arc structures using Elements AR. The area of each structure is calculated and the mean and SEM are reported for each nucleus, together with the number of puncta (N).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4596143&req=5

f1: Segmentation of nuclear substructures. A region of interest containing Arc-YFP puncta in a single neuronal nucleus is segmented to outline the Arc structures using Elements AR. The area of each structure is calculated and the mean and SEM are reported for each nucleus, together with the number of puncta (N).
Mentions: Fluorescence images were obtained using a motorized inverted wide-field epifluorescence microscope (Nikon Eclipse Ti-E), using 40× and 60× Plan-Apo oil objectives, with numerical apertures of 1.35 and 1.49, respectively. Motorized excitation and emission filter wheels (Ludl Electronics) fitted with a DAPI/CFP/YFP/DsRed quad filter set (#86010, Chroma) were used together with filter cubes for DAPI, CFP, YFP and TxRed, and Cy5 (Chroma) to select specific fluorescence signals. Z-stacks were obtained spanning the entire nucleus and out-of-focus fluorescence was removed using the AutoQuant 3D deconvolution algorithm (Media Cybernetics). Images were digitized using a cooled EM-CCD camera (iXon EM+ 885; Andor). Image acquisition was performed using NIS Elements AR 3.1 software (Nikon). NIS Elements analysis tools were used to outline the nuclei based on their DAPI images and to measure areas of the various nuclear substructures, as illustrated in Figure 1 for Arc-YFP puncta in the nucleus of a 20 DIV hippocampal neuron.

Bottom Line: Although Arc protein is found at synapses, it also localizes to the neuronal nucleus, where its function is less understood.Neuronal activity-induced expression of Arc (1) increases endogenous nuclear Tip60 puncta, (2) recruits Tip60 to PML bodies, and (3) increases histone acetylation of Tip60 substrate H4K12, a learning-induced chromatin modification.These mechanisms point to an epigenetic role for Arc in regulating memory consolidation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Program in Neuroscience and Behavioral Disorders, Duke-NUS Graduate Medical School , Singapore 169857.

ABSTRACT
Arc is an immediate-early gene whose genetic ablation selectively abrogates long-term memory, indicating a critical role in memory consolidation. Although Arc protein is found at synapses, it also localizes to the neuronal nucleus, where its function is less understood. Nuclear Arc forms a complex with the β-spectrin isoform βSpIVΣ5 and associates with PML bodies, sites of epigenetic regulation of gene expression. We report here a novel interaction between Arc and Tip60, a histone-acetyltransferase and subunit of a chromatin-remodelling complex, using biochemistry and super-resolution microscopy in primary rat hippocampal neurons. Arc and βSpIVΣ5 are recruited to nuclear Tip60 speckles, and the three proteins form a tight complex that localizes to nuclear perichromatin regions, sites of transcriptional activity. Neuronal activity-induced expression of Arc (1) increases endogenous nuclear Tip60 puncta, (2) recruits Tip60 to PML bodies, and (3) increases histone acetylation of Tip60 substrate H4K12, a learning-induced chromatin modification. These mechanisms point to an epigenetic role for Arc in regulating memory consolidation.

No MeSH data available.