PMID- 31752385
OWN - NLM
STAT- MEDLINE
DCOM- 20200707
LR  - 20210414
IS  - 2073-4409 (Electronic)
IS  - 2073-4409 (Linking)
VI  - 8
IP  - 11
DP  - 2019 Nov 19
TI  - Identification of Novel Adenylyl Cyclase 5 (AC5) Signaling Networks in D(1) and 
      D(2) Medium Spiny Neurons using Bimolecular Fluorescence Complementation 
      Screening.
LID - 10.3390/cells8111468 [doi]
LID - 1468
AB  - Adenylyl cyclase type 5 (AC5), as the principal isoform expressed in striatal 
      medium spiny neurons (MSNs), is essential for the integration of both stimulatory 
      and inhibitory midbrain signals that initiate from dopaminergic G protein-coupled 
      receptor (GPCR) activation. The spatial and temporal control of cAMP signaling is 
      dependent upon the composition of local regulatory protein networks. However, 
      there is little understanding of how adenylyl cyclase protein interaction 
      networks adapt to the multifarious pressures of integrating acute versus chronic 
      and inhibitory vs. stimulatory receptor signaling in striatal MSNs. Here, we 
      presented the development of a novel bimolecular fluorescence complementation 
      (BiFC)-based protein-protein interaction screening methodology to further 
      identify and characterize elements important for homeostatic control of 
      dopamine-modulated AC5 signaling in a neuronal model cell line and striatal MSNs. 
      We identified two novel AC5 modulators: the protein phosphatase 2A (PP2A) 
      catalytic subunit (PPP2CB) and the intracellular trafficking associated 
      protein-NSF (N-ethylmaleimide-sensitive factor) attachment protein alpha (NAPA). 
      The effects of genetic knockdown (KD) of each gene were evaluated in several 
      cellular models, including D(1)- and D(2)-dopamine receptor-expressing MSNs from 
      CAMPER mice. The knockdown of PPP2CB was associated with a reduction in acute and 
      sensitized adenylyl cyclase activity, implicating PP2A is an important and 
      persistent regulator of adenylyl cyclase activity. In contrast, the effects of 
      NAPA knockdown were more nuanced and appeared to involve an activity-dependent 
      protein interaction network. Taken together, these data represent a novel 
      screening method and workflow for the identification and validation of adenylyl 
      cyclase protein-protein interaction networks under diverse cAMP signaling 
      paradigms.
FAU - Doyle, Trevor B
AU  - Doyle TB
AD  - Medicinal Chemistry & Molecular Pharmacology, Purdue University, West Lafayette, 
      IN 47906, USA.
FAU - Muntean, Brian S
AU  - Muntean BS
AD  - Department of Neuroscience, The Scripps Research Institute, Jupiter, FL 33458, 
      USA.
FAU - Ejendal, Karin F
AU  - Ejendal KF
AUID- ORCID: 0000-0003-2099-0550
AD  - Medicinal Chemistry & Molecular Pharmacology, Purdue University, West Lafayette, 
      IN 47906, USA.
FAU - Hayes, Michael P
AU  - Hayes MP
AD  - Medicinal Chemistry & Molecular Pharmacology, Purdue University, West Lafayette, 
      IN 47906, USA.
FAU - Soto-Velasquez, Monica
AU  - Soto-Velasquez M
AD  - Medicinal Chemistry & Molecular Pharmacology, Purdue University, West Lafayette, 
      IN 47906, USA.
FAU - Martemyanov, Kirill A
AU  - Martemyanov KA
AD  - Department of Neuroscience, The Scripps Research Institute, Jupiter, FL 33458, 
      USA.
FAU - Dessauer, Carmen W
AU  - Dessauer CW
AUID- ORCID: 0000-0003-1210-4280
AD  - Department of Integrative Biology & Pharmacology, McGovern Medical School, 
      University of Texas Health Science Center, Houston, TX 77030, USA.
FAU - Hu, Chang-Deng
AU  - Hu CD
AD  - Medicinal Chemistry & Molecular Pharmacology, Purdue University, West Lafayette, 
      IN 47906, USA.
AD  - Purdue Institute of Drug Discovery, Purdue University, West Lafayette, IN 47906, 
      USA.
FAU - Watts, Val J
AU  - Watts VJ
AUID- ORCID: 0000-0003-1581-2936
AD  - Medicinal Chemistry & Molecular Pharmacology, Purdue University, West Lafayette, 
      IN 47906, USA.
AD  - Purdue Institute of Drug Discovery, Purdue University, West Lafayette, IN 47906, 
      USA.
AD  - Purdue Institute for Integrative Neuroscience, Purdue University, West Lafayette, 
      IN 47906, USA.
LA  - eng
GR  - R21 MH096927/MH/NIMH NIH HHS/United States
GR  - K02 DA026405/DA/NIDA NIH HHS/United States
GR  - R01 DA036596/DA/NIDA NIH HHS/United States
GR  - R21 MH101673/MH/NIMH NIH HHS/United States
GR  - R33 MH101673/MH/NIMH NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20191119
PL  - Switzerland
TA  - Cells
JT  - Cells
JID - 101600052
RN  - 0 (Carrier Proteins)
RN  - E0399OZS9N (Cyclic AMP)
RN  - EC 4.6.1.1 (Adenylyl Cyclases)
RN  - EC 4.6.1.1 (adenylyl cyclase type V)
RN  - VTD58H1Z2X (Dopamine)
SB  - IM
MH  - Adenylyl Cyclases/*metabolism
MH  - Animals
MH  - CRISPR-Cas Systems
MH  - Carrier Proteins/metabolism
MH  - Cyclic AMP/metabolism
MH  - Dopamine/metabolism
MH  - Drug Discovery
MH  - HEK293 Cells
MH  - Humans
MH  - Mice
MH  - Models, Biological
MH  - Neurons/drug effects/*metabolism
MH  - Protein Binding
MH  - *Signal Transduction/drug effects
PMC - PMC6912275
OTO - NOTNLM
OT  - AC5
OT  - BiFC
OT  - NAPA
OT  - PP2A
OT  - adenylyl cyclase
OT  - cAMP
OT  - dopamine
OT  - striatum
COIS- The authors declare no conflict of interest. The funders had no role in the 
      design of the study; in the collection, analyses, or interpretation of data; in 
      the writing of the manuscript, or in the decision to publish the results.
EDAT- 2019/11/23 06:00
MHDA- 2020/07/08 06:00
PMCR- 2019/11/01
CRDT- 2019/11/23 06:00
PHST- 2019/10/24 00:00 [received]
PHST- 2019/11/15 00:00 [revised]
PHST- 2019/11/15 00:00 [accepted]
PHST- 2019/11/23 06:00 [entrez]
PHST- 2019/11/23 06:00 [pubmed]
PHST- 2020/07/08 06:00 [medline]
PHST- 2019/11/01 00:00 [pmc-release]
AID - cells8111468 [pii]
AID - cells-08-01468 [pii]
AID - 10.3390/cells8111468 [doi]
PST - epublish
SO  - Cells. 2019 Nov 19;8(11):1468. doi: 10.3390/cells8111468.