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Distinct G protein – coupled receptor recycling pathways allow spatial control of downstream G protein signaling

View Article: PubMed Central - HTML - PubMed

ABSTRACT

GPCRs can activate different programs of gene expression from the plasma membrane and the endosome. Bowman et al. show that signaling by endosomal β-2 adrenergic receptors occurs at the microdomains that GPCRs use for sequence-dependent recycling.

No MeSH data available.


Related in: MedlinePlus

B2AR localization to ASRT domains correlates with expression of endosomal cAMP-specific genes. (A–C) Tukey box plots of fold change in PCK1 (A), CGA (B), and NR4A1 (C) expression across multiple experimental replicates. Iso increased PCK1 expression. Dynasore, Dyngo-4a, dynamin-K44A, or latA prevented the iso-induced increase in PCK1 expression. n = 11 for iso, iso + dynasore, and iso + latA, and n = 6 for all others. (D–F) Fold change in PCK1 (D), CGA (E), and NR4A1 (F) expression with iso for B2AR, SS>AA, and SS>DD. Iso increased PCK1 expression for B2AR and SS>DD, but not for SS>AA. n = 14 for B2AR and SS>AA, and n = 6 for SS>DD. Dashed lines throughout indicate the location of value 1 on the y axis. In all cases, p-values in relation to iso treatment alone are shown.
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fig5: B2AR localization to ASRT domains correlates with expression of endosomal cAMP-specific genes. (A–C) Tukey box plots of fold change in PCK1 (A), CGA (B), and NR4A1 (C) expression across multiple experimental replicates. Iso increased PCK1 expression. Dynasore, Dyngo-4a, dynamin-K44A, or latA prevented the iso-induced increase in PCK1 expression. n = 11 for iso, iso + dynasore, and iso + latA, and n = 6 for all others. (D–F) Fold change in PCK1 (D), CGA (E), and NR4A1 (F) expression with iso for B2AR, SS>AA, and SS>DD. Iso increased PCK1 expression for B2AR and SS>DD, but not for SS>AA. n = 14 for B2AR and SS>AA, and n = 6 for SS>DD. Dashed lines throughout indicate the location of value 1 on the y axis. In all cases, p-values in relation to iso treatment alone are shown.

Mentions: To test whether endosome-specific cAMP responses were regulated by PKA-mediated B2AR relocation, we used reverse transcription followed by quantitative real-time PCR to measure the expression of three genes—PCK1 (phosphoenolpyruvate carboxykinase 1), CGA (glycoprotein hormones, α-polypeptide), and NR4A1 (nuclear receptor subfamily 4 group A member 1)—that are induced specifically by endosomal cAMP (Tsvetanova and von Zastrow, 2014). We isolated RNA from HEK 293 cells stably expressing wild-type B2AR or SS>AA and measured the expression of PCK1 as well as the reference gene, β-tubulin (TUBB). Dose–response curve analysis showed that TUBB expression changed in a similar concentration-dependent manner with and without iso treatment (Fig. S3, D and E), making it a suitable gene to normalize the expression of iso-dependent genes across treatments. When normalized to TUBB expression, PCK1, CGA, and NR4A1 expression levels increased by approximately five-, six-, and eightfold upon B2AR activation by iso (Fig. 5, A–C).


Distinct G protein – coupled receptor recycling pathways allow spatial control of downstream G protein signaling
B2AR localization to ASRT domains correlates with expression of endosomal cAMP-specific genes. (A–C) Tukey box plots of fold change in PCK1 (A), CGA (B), and NR4A1 (C) expression across multiple experimental replicates. Iso increased PCK1 expression. Dynasore, Dyngo-4a, dynamin-K44A, or latA prevented the iso-induced increase in PCK1 expression. n = 11 for iso, iso + dynasore, and iso + latA, and n = 6 for all others. (D–F) Fold change in PCK1 (D), CGA (E), and NR4A1 (F) expression with iso for B2AR, SS>AA, and SS>DD. Iso increased PCK1 expression for B2AR and SS>DD, but not for SS>AA. n = 14 for B2AR and SS>AA, and n = 6 for SS>DD. Dashed lines throughout indicate the location of value 1 on the y axis. In all cases, p-values in relation to iso treatment alone are shown.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC5037407&req=5

fig5: B2AR localization to ASRT domains correlates with expression of endosomal cAMP-specific genes. (A–C) Tukey box plots of fold change in PCK1 (A), CGA (B), and NR4A1 (C) expression across multiple experimental replicates. Iso increased PCK1 expression. Dynasore, Dyngo-4a, dynamin-K44A, or latA prevented the iso-induced increase in PCK1 expression. n = 11 for iso, iso + dynasore, and iso + latA, and n = 6 for all others. (D–F) Fold change in PCK1 (D), CGA (E), and NR4A1 (F) expression with iso for B2AR, SS>AA, and SS>DD. Iso increased PCK1 expression for B2AR and SS>DD, but not for SS>AA. n = 14 for B2AR and SS>AA, and n = 6 for SS>DD. Dashed lines throughout indicate the location of value 1 on the y axis. In all cases, p-values in relation to iso treatment alone are shown.
Mentions: To test whether endosome-specific cAMP responses were regulated by PKA-mediated B2AR relocation, we used reverse transcription followed by quantitative real-time PCR to measure the expression of three genes—PCK1 (phosphoenolpyruvate carboxykinase 1), CGA (glycoprotein hormones, α-polypeptide), and NR4A1 (nuclear receptor subfamily 4 group A member 1)—that are induced specifically by endosomal cAMP (Tsvetanova and von Zastrow, 2014). We isolated RNA from HEK 293 cells stably expressing wild-type B2AR or SS>AA and measured the expression of PCK1 as well as the reference gene, β-tubulin (TUBB). Dose–response curve analysis showed that TUBB expression changed in a similar concentration-dependent manner with and without iso treatment (Fig. S3, D and E), making it a suitable gene to normalize the expression of iso-dependent genes across treatments. When normalized to TUBB expression, PCK1, CGA, and NR4A1 expression levels increased by approximately five-, six-, and eightfold upon B2AR activation by iso (Fig. 5, A–C).

View Article: PubMed Central - HTML - PubMed

ABSTRACT

GPCRs can activate different programs of gene expression from the plasma membrane and the endosome. Bowman et al. show that signaling by endosomal β-2 adrenergic receptors occurs at the microdomains that GPCRs use for sequence-dependent recycling.

No MeSH data available.


Related in: MedlinePlus