SPCA2 Regulates Orai1 Trafficking and Store Independent Ca2+ Entry in a Model of Lactation

Brandie M. Cross, Anniesha Hack, Timothy A. Reinhardt, Rajini Rao

Research output: Contribution to journalArticle

Abstract

An unconventional interaction between SPCA2, an isoform of the Golgi secretory pathway Ca2+-ATPase, and the Ca2+ influx channel Orai1, has previously been shown to contribute to elevated Ca2+ influx in breast cancer derived cells. In order to investigate the physiological role of this interaction, we examined expression and localization of SPCA2 and Orai1 in mouse lactating mammary glands. We observed co-induction and co-immunoprecipitation of both proteins, and isoform-specific differences in the localization of SPCA1 and SPCA2. Three-dimensional cultures of normal mouse mammary epithelial cells were established using lactogenic hormones and basement membrane. The mammospheres displayed elevated Ca2+ influx by store independent mechanisms, consistent with upregulation of both SPCA2 and Orai1. Knockdown of either SPCA2 or Orai1 severely depleted Ca2+ influx and interfered with mammosphere differentiation. We show that SPCA2 is required for plasma membrane trafficking of Orai1 in mouse mammary epithelial cells and that this function can be replaced, at least in part, by a membrane-anchored C-terminal domain of SPCA2. These findings clearly show that SPCA2 and Orai1 function together to regulate Store-independent Ca2+ entry (SICE), which mediates the massive basolateral Ca2+ influx into mammary epithelia to support the large calcium transport requirements for milk secretion.

Original languageEnglish (US)
Article numbere67348
JournalPLoS One
Volume8
Issue number6
DOIs
StatePublished - Jun 28 2013

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Lactation
Protein Isoforms
Breast
lactation
calcium
Calcium-Transporting ATPases
Cell membranes
Epithelial Cells
breasts
Secretory Pathway
Hormones
Human Mammary Glands
Calcium
Membranes
Immunoprecipitation
Basement Membrane
Milk
Up-Regulation
Epithelium
Cell Membrane

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

SPCA2 Regulates Orai1 Trafficking and Store Independent Ca2+ Entry in a Model of Lactation. / Cross, Brandie M.; Hack, Anniesha; Reinhardt, Timothy A.; Rao, Rajini.

In: PLoS One, Vol. 8, No. 6, e67348, 28.06.2013.

Research output: Contribution to journalArticle

Cross, Brandie M. ; Hack, Anniesha ; Reinhardt, Timothy A. ; Rao, Rajini. / SPCA2 Regulates Orai1 Trafficking and Store Independent Ca2+ Entry in a Model of Lactation. In: PLoS One. 2013 ; Vol. 8, No. 6.
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