Role of membrane microdomains in compartmentation of cAMP signaling.
Spatially restricting cAMP production to discrete subcellular locations permits selective regulation of specific functional responses. But exactly where and how cAMP signaling is confined is not fully understood. Different receptors and adenylyl cyclase isoforms responsible for cAMP production are n...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2014-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3994114?pdf=render |
| _version_ | 1818280815438594048 |
|---|---|
| author | Shailesh R Agarwal Pei-Chi Yang Monica Rice Cherie A Singer Viacheslav O Nikolaev Martin J Lohse Colleen E Clancy Robert D Harvey |
| author_facet | Shailesh R Agarwal Pei-Chi Yang Monica Rice Cherie A Singer Viacheslav O Nikolaev Martin J Lohse Colleen E Clancy Robert D Harvey |
| author_sort | Shailesh R Agarwal |
| collection | DOAJ |
| description | Spatially restricting cAMP production to discrete subcellular locations permits selective regulation of specific functional responses. But exactly where and how cAMP signaling is confined is not fully understood. Different receptors and adenylyl cyclase isoforms responsible for cAMP production are not uniformly distributed between lipid raft and non-lipid raft domains of the plasma membrane. We sought to determine the role that these membrane domains play in organizing cAMP responses in HEK293 cells. The freely diffusible FRET-based biosensor Epac2-camps was used to measure global cAMP responses, while versions of the probe targeted to lipid raft (Epac2-MyrPalm) and non-raft (Epac2-CAAX) domains were used to monitor local cAMP production near the plasma membrane. Disruption of lipid rafts by cholesterol depletion selectively altered cAMP responses produced by raft-associated receptors. The results indicate that receptors associated with lipid raft as well as non-lipid raft domains can contribute to global cAMP responses. In addition, basal cAMP activity was found to be significantly higher in non-raft domains. This was supported by the fact that pharmacologic inhibition of adenylyl cyclase activity reduced basal cAMP activity detected by Epac2-CAAX but not Epac2-MyrPalm or Epac2-camps. Responses detected by Epac2-CAAX were also more sensitive to direct stimulation of adenylyl cyclase activity, but less sensitive to inhibition of phosphodiesterase activity. Quantitative modeling was used to demonstrate that differences in adenylyl cyclase and phosphodiesterase activities are necessary but not sufficient to explain compartmentation of cAMP associated with different microdomains of the plasma membrane. |
| first_indexed | 2024-12-12T23:55:13Z |
| format | Article |
| id | doaj.art-8e03eb1e909b4ba8ac469ca509655384 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-12T23:55:13Z |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-8e03eb1e909b4ba8ac469ca5096553842022-12-22T00:06:35ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0194e9583510.1371/journal.pone.0095835Role of membrane microdomains in compartmentation of cAMP signaling.Shailesh R AgarwalPei-Chi YangMonica RiceCherie A SingerViacheslav O NikolaevMartin J LohseColleen E ClancyRobert D HarveySpatially restricting cAMP production to discrete subcellular locations permits selective regulation of specific functional responses. But exactly where and how cAMP signaling is confined is not fully understood. Different receptors and adenylyl cyclase isoforms responsible for cAMP production are not uniformly distributed between lipid raft and non-lipid raft domains of the plasma membrane. We sought to determine the role that these membrane domains play in organizing cAMP responses in HEK293 cells. The freely diffusible FRET-based biosensor Epac2-camps was used to measure global cAMP responses, while versions of the probe targeted to lipid raft (Epac2-MyrPalm) and non-raft (Epac2-CAAX) domains were used to monitor local cAMP production near the plasma membrane. Disruption of lipid rafts by cholesterol depletion selectively altered cAMP responses produced by raft-associated receptors. The results indicate that receptors associated with lipid raft as well as non-lipid raft domains can contribute to global cAMP responses. In addition, basal cAMP activity was found to be significantly higher in non-raft domains. This was supported by the fact that pharmacologic inhibition of adenylyl cyclase activity reduced basal cAMP activity detected by Epac2-CAAX but not Epac2-MyrPalm or Epac2-camps. Responses detected by Epac2-CAAX were also more sensitive to direct stimulation of adenylyl cyclase activity, but less sensitive to inhibition of phosphodiesterase activity. Quantitative modeling was used to demonstrate that differences in adenylyl cyclase and phosphodiesterase activities are necessary but not sufficient to explain compartmentation of cAMP associated with different microdomains of the plasma membrane.http://europepmc.org/articles/PMC3994114?pdf=render |
| spellingShingle | Shailesh R Agarwal Pei-Chi Yang Monica Rice Cherie A Singer Viacheslav O Nikolaev Martin J Lohse Colleen E Clancy Robert D Harvey Role of membrane microdomains in compartmentation of cAMP signaling. PLoS ONE |
| title | Role of membrane microdomains in compartmentation of cAMP signaling. |
| title_full | Role of membrane microdomains in compartmentation of cAMP signaling. |
| title_fullStr | Role of membrane microdomains in compartmentation of cAMP signaling. |
| title_full_unstemmed | Role of membrane microdomains in compartmentation of cAMP signaling. |
| title_short | Role of membrane microdomains in compartmentation of cAMP signaling. |
| title_sort | role of membrane microdomains in compartmentation of camp signaling |
| url | http://europepmc.org/articles/PMC3994114?pdf=render |
| work_keys_str_mv | AT shaileshragarwal roleofmembranemicrodomainsincompartmentationofcampsignaling AT peichiyang roleofmembranemicrodomainsincompartmentationofcampsignaling AT monicarice roleofmembranemicrodomainsincompartmentationofcampsignaling AT cherieasinger roleofmembranemicrodomainsincompartmentationofcampsignaling AT viacheslavonikolaev roleofmembranemicrodomainsincompartmentationofcampsignaling AT martinjlohse roleofmembranemicrodomainsincompartmentationofcampsignaling AT colleeneclancy roleofmembranemicrodomainsincompartmentationofcampsignaling AT robertdharvey roleofmembranemicrodomainsincompartmentationofcampsignaling |