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Characterization and Transcriptional Activation of the Immediate Early Gene ARC During a Neural Correlate of Classical Conditioning

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Abstract

Plasticity and learning genes require regulatory mechanisms that have the flexibility to respond to a variety of sensory stimuli to generate adaptive behavioral responses. The immediate early gene (IEG) activity–regulated cytoskeleton-associated protein (ARC) is rapidly induced not only by neuronal stimulation but also during a variety of learning tasks. How ARC is regulated in response to complex stimuli during associative learning remains to be fully detailed. Here, we characterized the structure of the ARC gene in the pond turtle and mechanisms of its transcriptional activation during a neural correlate of eyeblink classical conditioning. The tARC gene is regulated in part by the presence of paused polymerase (RNAPII) that is poised at the promoter for rapid gene induction. Conditioning induces permissive chromatin modifications in the tARC promoter that allows binding by the transcription factor cAMP response element–binding protein (CREB) within 5 min of training. During learning acquisition, the pausing factor negative elongation factor (NELF) dissociates from the promoter thereby releasing RNAPII for active transcription. Data additionally suggest that the DNA insulator protein CCCTC–binding factor (CTCF) is required for transcription by mediating a learning-induced interaction of the ARC promoter with an enhancer element. Our study suggests that the learning-inducible IEG tARC utilizes both paused RNAPII and rapid chromatin modifications that allow for dynamic gene responsiveness required when an organism is presented with a variety of environmental stimuli.

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Funding

J.K was supported by institutional departmental funds.

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Authors and Affiliations

  1. Neuroscience Group, Division of Basic Biomedical Sciences, University of South Dakota Sanford School of Medicine, 414 E. Clark St, Vermillion, SD, 57069, USA

    Zhaoqing Zheng, Ganesh Ambigapathy & Joyce Keifer

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  1. Zhaoqing Zheng
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  2. Ganesh Ambigapathy
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  3. Joyce Keifer
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Corresponding author

Correspondence to Joyce Keifer.

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All experiments involving the use of animals were performed in accordance with the guidelines National Institutes of Health and were approved by the University of South Dakota Institutional Animal Care and Use Committee.

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Supplemental Fig. 1

The cDNA sequence of the 5′ flanking region and exons I-III of tARC for T. scripta elegans (GenBank: MG851989.1). Asterisks mark the transcription start sites. The protein coding sequence is shown in italics. The start and stop codons are indicated as are some of the notable transcription factor binding sites and polyadenylation sites. Primers for RLM-RACE are also shown. (PNG 3180 kb)

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Zheng, Z., Ambigapathy, G. & Keifer, J. Characterization and Transcriptional Activation of the Immediate Early Gene ARC During a Neural Correlate of Classical Conditioning. J Mol Neurosci 69, 380–390 (2019). https://doi.org/10.1007/s12031-019-01367-z

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  • DOI: https://doi.org/10.1007/s12031-019-01367-z

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