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J. Biol. Chem., Vol. 277, Issue 27, 24442-24452, July 5, 2002

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Three Novel Sarco/endoplasmic Reticulum Ca²⁺-ATPase (SERCA) 3 Isoforms
EXPRESSION, REGULATION, AND FUNCTION OF THE MEMBERS OF THE SERCA3 FAMILY^*

Virginie Martin^§, Raymonde Bredoux, Elisabeth Corvazier, Roosje van Gorp, Tünde Kovàcs^¶, Pascal Gélébart, and Jocelyne Enouf^**

From  INSERM U348, IFR6 Circulation Lariboisière, Hôpital Lariboisière, 8 Rue Guy Patin, 75475 Paris Cedex 10, France and the ^¶ National Medical Center, Institute of Haematology and Immunology, H-1113 Budapest, Hungary

Sarco/endoplasmic reticulum Ca²⁺-ATPases (SERCAs) pump Ca²⁺ into the endoplasmic reticulum. Recently, three human SERCA3 (h3a-c) proteins and a previously unknown rat SERCA3 (r3b/c) mRNA have been described. Here, we (i) document two novel human SERCA3 splice variants h3d and h3e, (ii) provide data for the expression and mechanisms regulating the expression of all known SERCA3 variants (r3a, r3b/c, and h3a-e), and (iii) show functional characteristics of the SERCA3 isoforms. h3d and h3e are issued from the insertion of an additional penultimate exon 22 resulting in different carboxyl termini for these variants. Distinct distribution patterns of the SERCA3 gene products were observed in a series of cell lines of hematopoietic, epithelial, embryonic origin, and several cancerous types, as well as in panels of rat and human tissues. Hypertension and protein kinase C, calcineurin, or retinoic acid receptor signaling pathways were found to differently control rat and human splice variant expression, respectively. Stable overexpression of each variant was performed in human embryonic kidney 293 cells, and the SERCA3 isoforms were fully characterized. All SERCA3 isoforms were found to pump Ca²⁺ with similar affinities. However, they modulated the cytosolic Ca²⁺ concentration ([Ca²⁺]_c) and the endoplasmic reticulum Ca²⁺ content ([Ca²⁺]_er) in different manners. A newly generated polyclonal antibody and a pan-SERCA3 antibody proved the endogenous expression of the three novel SERCA3 proteins, h3d, h3e, and r3b/c. All these data suggest that the SERCA3 gene products have a more widespread role in cellular Ca²⁺ signaling than previously appreciated.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AF458228-AF458230.

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