Dataset Information

Phosphate (P_i)-regulated heterodimerization of the high-affinity sodium-dependent P_i transporters PiT1/Slc20a1 and PiT2/Slc20a2 underlies extracellular P_i sensing independently of P_i uptake.

ABSTRACT: Extracellular phosphate (P_i) can act as a signaling molecule that directly alters gene expression and cellular physiology. The ability of cells or organisms to detect changes in extracellular P_i levels implies the existence of a P_i-sensing mechanism that signals to the body or individual cell. However, unlike in prokaryotes, yeasts, and plants, the molecular players involved in P_i sensing in mammals remain unknown. In this study, we investigated the involvement of the high-affinity, sodium-dependent P_i transporters PiT1 and PiT2 in mediating P_i signaling in skeletal cells. We found that deletion of PiT1 or PiT2 blunted the P_i-dependent ERK1/2-mediated phosphorylation and subsequent gene up-regulation of the mineralization inhibitors matrix Gla protein and osteopontin. This result suggested that both PiTs are necessary for P_i signaling. Moreover, the ERK1/2 phosphorylation could be rescued by overexpressing P_i transport-deficient PiT mutants. Using cross-linking and bioluminescence resonance energy transfer approaches, we found that PiT1 and PiT2 form high-abundance homodimers and P_i-regulated low-abundance heterodimers. Interestingly, in the absence of sodium-dependent P_i transport activity, the PiT1-PiT2 heterodimerization was still regulated by extracellular P_i levels. Of note, when two putative P_i-binding residues, Ser-128 (in PiT1) and Ser-113 (in PiT2), were substituted with alanine, the PiT1-PiT2 heterodimerization was no longer regulated by extracellular P_i These observations suggested that P_i binding rather than P_i uptake may be the key factor in mediating P_i signaling through the PiT proteins. Taken together, these results demonstrate that P_i-regulated PiT1-PiT2 heterodimerization mediates P_i sensing independently of P_i uptake.

SUBMITTER: Bon N

PROVIDER: S-EPMC5808770 | biostudies-literature | 2018 Feb

REPOSITORIES: biostudies-literature

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Phosphate (Pi)-regulated heterodimerization of the high-affinity sodium-dependent Pi transporters PiT1/Slc20a1 and PiT2/Slc20a2 underlies extracellular Pi sensing independently of Pi uptake.

Bon Nina N Couasnay Greig G Bourgine Annabelle A Sourice Sophie S Beck-Cormier Sarah S Guicheux Jérôme J Beck Laurent L

The Journal of biological chemistry 20171212 6

Extracellular phosphate (Pi) can act as a signaling molecule that directly alters gene expression and cellular physiology. The ability of cells or organisms to detect changes in extracellular Pi levels implies the existence of a Pi-sensing mechanism that signals to the body or individual cell. However, unlike in prokaryotes, yeasts, and plants, the molecular players involved in Pi sensing in mammals remain unknown. In this study, we investigated the in ...[more]

PMID: 29233890

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Project description:Subgroup B feline leukemia viruses (FeLV-Bs) evolve from subgroup A FeLV (FeLV-A) by recombining with portions of endogenous FeLV envelope sequences in the cat genome. The replication properties of FeLV-B are distinct from those of FeLV-A; FeLV-B infects many nonfeline cell lines and recognizes the human Pit1 (HuPit1) receptor, whereas FeLV-A infects primarily feline cells, using a distinct but as yet undefined receptor. Here, we demonstrate that some FeLV-Bs can also use human Pit2 (HuPit2) and hamster Pit2 (HaPit2) for entry. By making viruses that contain chimeric surface (SU) envelope proteins from FeLV-A and FeLV-B, and testing their infectivity, we have defined genetic determinants that confer host range and specific receptor recognition. HuPit1 receptor recognition determinants localize to the N-terminal region of the FeLV-B SU, amino acids 83 to 116, encompassing the N-terminal portion of variable region A (VRA). While this 34-amino-acid domain of the FeLV-B VRA is sufficient for infection of some cells (feline, canine, and human), amino acids 146 to 249 of FeLV-B, which include variable region B (VRB), were required for efficient infection in other cell types (hamster, bovine, and rat). Chimeras encoding FeLV-B VRA and VRB also infected cells expressing HaPit2 and HuPit2 receptors more efficiently than chimeras encoding only the VRA of FeLV-B, suggesting that VRB provides a secondary determinant that is both cell and receptor specific. However, viruses containing additional FeLV-B sequences in the C terminus of SU could not recognize HuPit2, implying that there is a determinant beyond VRB that negatively affects HuPit2 interactions. Thus, Pit2 recognition may drive selection for the generation of specific FeLV-B recombinants, offering an explanation for the two major classes of FeLV-B that have been observed in vivo. Furthermore, the finding that some FeLV-Bs can use both Pit1 and Pit2 may explain previous observations that FeLV-B and GALV, which primarily uses Pit1, display nonreciprocal interference on many cell types.

Dataset Information

Phosphate (P_i)-regulated heterodimerization of the high-affinity sodium-dependent P_i transporters PiT1/Slc20a1 and PiT2/Slc20a2 underlies extracellular P_i sensing independently of P_i uptake.

Publications

Phosphate (P<sub>i</sub>)-regulated heterodimerization of the high-affinity sodium-dependent P<sub>i</sub> transporters PiT1/Slc20a1 and PiT2/Slc20a2 underlies extracellular P<sub>i</sub> sensing independently of P<sub>i</sub> uptake.

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