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Permanent link (DOI): https://doi.org/10.7939/R33104

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Salt-Inducible Kinase 1 and Transducer or Regulated CREB Activity 2 in the Rat Pineal Gland Open Access

Descriptions

Other title
Subject/Keyword
pineal gland
rhythmic gene expression
protein kinase A
coactivator
cAMP response element binding protein
transducer of regulated CREB activity 2
salt-inducible kinase
arylalkylamine-N-acetyltransferase
cAMP
melatonin
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Kanyo, Richard
Supervisor and department
Ho, Anthony (Physiology)
Examining committee member and department
Karpinski, Ed (Physiology)
Gosgnach, Simon (Physiology)
Chik, Constance (Medicine)
Ezzat, Shereen (Medicine)
Morrish, Don (Medicine)
Department
Department of Physiology
Specialization

Date accepted
2013-11-07T13:53:36Z
Graduation date
2014-06
Degree
Doctor of Philosophy
Degree level
Doctoral
Abstract
In the rat pineal gland, the transcription of the melatonin-rhythm generating enzyme, arylalkyl-N-acetyltransferase (AA-NAT), is stimulated by norepinephrine (NE), acting through the cAMP → protein kinase A (PKA) → cAMP response element-binding protein (CREB) signalling pathway. Although the PKA-mediated phosphorylation of CREB is the main activation mechanism, the transcriptional activity of CREB can also be regulated by the salt-inducible kinase (SIK)/transducer of regulated CREB activity (TORC) pathway. Therefore, the objective of this study was to investigate the mechanisms involved in the regulation of SIK1 and TORC2 in the rat pineal gland and their roles in Aa-nat transcription. We found a marked nocturnal induction of Sik1 transcription in the rat pineal gland. In cultured pinealocytes, this induction is driven primarily by a β-adrenoreceptor/cAMP-dependent mechanism with a minor contribution from the α1-adrenoreceptor/Ca2+-pathway. Manipulating the level of Sik1 expression in pinealocytes shows that endogenous SIK1 can function as a transcription repressor of Aa-nat. Studies on TORC2 show that although the TORC2 protein is dephosphorylated in the rat pineal gland with the onset of darkness and in pinealocytes within 15 min of NE stimulation, this occurs without any changes in the Torc2 mRNA levels. The β-adrenoreceptor/cAMP-pathway and protein phosphatase 2A are involved in the NE-stimulated dephosphorylation of TORC2, which also result in its nuclear translocation. Elevating intracellular Ca2+ also induces dephosphorylation of TORC2 but has no effect on its nuclear translocation. As for the role of TORC2 on Aa-nat transcription, whereas overexpression of TORC2 has an enhancing effect on the NE-stimulated Aa-nat expression, knockdown of TORC2 only has a small inhibitory effect. Finally, knockdown of endogenous Sik1 has no effect on the phosphorylation status or cellular distribution of TORC2 under basal or stimulated conditions. We show that the β-adrenoreceptor/cAMP signalling mechanism is the dominant pathway in inducing Sik1 transcription and nuclear translocation of TORC2. However, the repressive effect of SIK1 on Aa-nat cannot be explained by regulating the cellular distribution of TORC2. Moreover, the phosphorylation status of TORC2 may not accurately reflect its activation and that TORC2 cellular distribution is not regulated solely by SIK1.
Language
English
DOI
doi:10.7939/R33104
Rights
Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
Citation for previous publication
Kanyo, R., D.M. Price, C.L. Chik, and A.K. Ho. 2009. Salt-inducible kinase 1 in the rat pinealocyte: adrenergic regulation and role in arylalkylamine N-acetyltransferase gene transcription. Endocrinology. 150:4221-4230.Kanyo, R., N. Amyotte, J. McTague, C.L. Chik, and A.K. Ho. 2011. Adrenergic regulation of the distribution of transducer of regulated cAMP-response element-binding protein (TORC2) in rat pinealocytes. Endocrinology. 152:3440-3450.McTague, J., N. Amyotte, R. Kanyo, M. Ferguson, C.L. Chik, and A.K. Ho. 2012. Different Signaling Mechanisms Are Involved in the Norepinephrine-Stimulated TORC1 and TORC2 Nuclear Translocation in Rat Pinealocytes. Endocrinology. 153:3839-49.

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