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Arrestins function in cAR1 GPCR-mediated signaling and cAR1 internalization in the development of Dictyostelium discoideum.

Cao X, Yan J, Shu S, Brzostowski JA, Jin T - Mol. Biol. Cell (2014)

Bottom Line: At the aggregation stage of Dictyostelium discoideum development, the chemoattractant cAMP is synthesized and released at 6-min intervals, directing cell migration.The adcB(-)C(-) cells also have a shorter period of transient ERK2 activity and precociously reactivate ERK2 in response to cAMP stimulation.In addition, ligand-induced cAR1 internalization is compromised in adcB(-)C(-) cells, suggesting that arrestins are involved in elimination of high-affinity cAR1 receptors from cell surface after the aggregation stage of multicellular development.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Institute of Immunology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.

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Arrestin-like proteins function in the development of D. discoideum. (A) Development of arrestin  cells on bacterial lawns. Wild-type, adcB−, adcC−, adcB−C−, and adcB−C− cells expressing AdcB-YFP or AdcC-YFP were grown in association with K. aerogenes at 22°C. Photographs were taken after 5 d. (B) The number of mounds in plaques formed by cells was counted and graphed. Means (n = 4–6) and SDs are shown. Statistical significance was assessed by t test, *p < 0.05 and **p < 0.01. (C) Development of arrestin  cells on nonnutrient agar. Arrestin- cells were plated on nonnutrient agar to initiate starvation and the developmental program. Images were captured at the indicated times to show aggregation (5 h), slug (10 h), and fruiting body (24 h) stages.
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Figure 1: Arrestin-like proteins function in the development of D. discoideum. (A) Development of arrestin cells on bacterial lawns. Wild-type, adcB−, adcC−, adcB−C−, and adcB−C− cells expressing AdcB-YFP or AdcC-YFP were grown in association with K. aerogenes at 22°C. Photographs were taken after 5 d. (B) The number of mounds in plaques formed by cells was counted and graphed. Means (n = 4–6) and SDs are shown. Statistical significance was assessed by t test, *p < 0.05 and **p < 0.01. (C) Development of arrestin cells on nonnutrient agar. Arrestin- cells were plated on nonnutrient agar to initiate starvation and the developmental program. Images were captured at the indicated times to show aggregation (5 h), slug (10 h), and fruiting body (24 h) stages.

Mentions: To reveal the potential roles of these genes in the development of D. discoideum, we analyzed the developmental phenotypes of adcB−, adcC−, and adcB −C− cells on bacterial lawns (Figure 1A and Supplemental Figure S3) and on nonnutrient agar (Figure 1C). Whereas adcB− and adcC− cells displayed a wild-type-like phenotype, adcB−C− cells were unable to form a normal number of mounds in each plaque (Figure 1, A and B) and failed to aggregate by 5 h, but they eventually formed smaller aggregates at 10 h on nonnutrient agar (Figure 1C). The developmental phenotype was partially rescued by expressing either yellow fluorescent protein (YFP)–tagged AdcB or AdcC (Figure 1, A and B), indicating that either AdcB or AdcC is required for proper development of D. discoideum and that both AdcB-YFP and AdcC-YFP are functional.


Arrestins function in cAR1 GPCR-mediated signaling and cAR1 internalization in the development of Dictyostelium discoideum.

Cao X, Yan J, Shu S, Brzostowski JA, Jin T - Mol. Biol. Cell (2014)

Arrestin-like proteins function in the development of D. discoideum. (A) Development of arrestin  cells on bacterial lawns. Wild-type, adcB−, adcC−, adcB−C−, and adcB−C− cells expressing AdcB-YFP or AdcC-YFP were grown in association with K. aerogenes at 22°C. Photographs were taken after 5 d. (B) The number of mounds in plaques formed by cells was counted and graphed. Means (n = 4–6) and SDs are shown. Statistical significance was assessed by t test, *p < 0.05 and **p < 0.01. (C) Development of arrestin  cells on nonnutrient agar. Arrestin- cells were plated on nonnutrient agar to initiate starvation and the developmental program. Images were captured at the indicated times to show aggregation (5 h), slug (10 h), and fruiting body (24 h) stages.
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Related In: Results  -  Collection

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Figure 1: Arrestin-like proteins function in the development of D. discoideum. (A) Development of arrestin cells on bacterial lawns. Wild-type, adcB−, adcC−, adcB−C−, and adcB−C− cells expressing AdcB-YFP or AdcC-YFP were grown in association with K. aerogenes at 22°C. Photographs were taken after 5 d. (B) The number of mounds in plaques formed by cells was counted and graphed. Means (n = 4–6) and SDs are shown. Statistical significance was assessed by t test, *p < 0.05 and **p < 0.01. (C) Development of arrestin cells on nonnutrient agar. Arrestin- cells were plated on nonnutrient agar to initiate starvation and the developmental program. Images were captured at the indicated times to show aggregation (5 h), slug (10 h), and fruiting body (24 h) stages.
Mentions: To reveal the potential roles of these genes in the development of D. discoideum, we analyzed the developmental phenotypes of adcB−, adcC−, and adcB −C− cells on bacterial lawns (Figure 1A and Supplemental Figure S3) and on nonnutrient agar (Figure 1C). Whereas adcB− and adcC− cells displayed a wild-type-like phenotype, adcB−C− cells were unable to form a normal number of mounds in each plaque (Figure 1, A and B) and failed to aggregate by 5 h, but they eventually formed smaller aggregates at 10 h on nonnutrient agar (Figure 1C). The developmental phenotype was partially rescued by expressing either yellow fluorescent protein (YFP)–tagged AdcB or AdcC (Figure 1, A and B), indicating that either AdcB or AdcC is required for proper development of D. discoideum and that both AdcB-YFP and AdcC-YFP are functional.

Bottom Line: At the aggregation stage of Dictyostelium discoideum development, the chemoattractant cAMP is synthesized and released at 6-min intervals, directing cell migration.The adcB(-)C(-) cells also have a shorter period of transient ERK2 activity and precociously reactivate ERK2 in response to cAMP stimulation.In addition, ligand-induced cAR1 internalization is compromised in adcB(-)C(-) cells, suggesting that arrestins are involved in elimination of high-affinity cAR1 receptors from cell surface after the aggregation stage of multicellular development.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Institute of Immunology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.

Show MeSH
Related in: MedlinePlus