Transmembrane nine proteins in yeast and Arabidopsis affect cellular metal contents without changing vacuolar morphology

TY - JOUR

T1 - Transmembrane nine proteins in yeast and Arabidopsis affect cellular metal contents without changing vacuolar morphology

AU - Hegelund, Josefine Nymark

AU - Jahn, Thomas Paul

AU - Bækgaard, Lone

AU - Palmgren, Michael

AU - Schjørring, Jan Kofod

PY - 2010/12

Y1 - 2010/12

N2 - Transmembrane nine (TM9) proteins are localized in the secretory pathway of eukaryotic cells and are involved in cell adhesion and phagocytosis. The mechanism by which TM9 proteins operate is, however, not well understood. Here we have utilized elemental profiling by inductively coupled plasma mass spectrometry (ICP-MS) to further investigate the physiological function of TM9 proteins. Cellular copper contents in Saccharomyces cerevisiae varied depending on the presence of TM9 homologues from both yeast and Arabidopsis thaliana. A yeast tmn1-3 triple mutant lacking all three yeast endogenous TMNs showed altered metal homeostasis with a reduction in the cellular Cu contents to 25% of that in the wild-type. Conversely, when TMN1 was overexpressed in yeast, cellular Cu concentrations were more than doubled. Both Tmn1p-GFP and Tmn2p-GFP fusion proteins localized to the tonoplast. Yeast vacuolar biogenesis was not affected by the lack or presence of TM9 proteins neither in the tmn1-3 triple mutant nor in TM9 overexpressing strains. Heterologous expression in yeast of AtTMN7, a TM9 homologue from Arabidopsis, affected Cu homeostasis similar to the overexpression of TMN1. In Arabidopsis, the two TM9 homologues AtTMN1 and AtTMN7 were ubiquitously expressed. AtTMN7 promoter constructs driving the expression of GFP showed elevated expression of AtTMN7 in the root elongation zone. It is concluded that TM9 homologues from S. cerevisiae and A. thaliana have the ability to affect the intracellular Cu balance. Tmn1p and Tmn2p operate from the yeast vacuolar membrane without influencing vacuolar biogenesis. A new physiological function of the TM9 family coupled to vacuolar Cu homeostasis is proposed.

AB - Transmembrane nine (TM9) proteins are localized in the secretory pathway of eukaryotic cells and are involved in cell adhesion and phagocytosis. The mechanism by which TM9 proteins operate is, however, not well understood. Here we have utilized elemental profiling by inductively coupled plasma mass spectrometry (ICP-MS) to further investigate the physiological function of TM9 proteins. Cellular copper contents in Saccharomyces cerevisiae varied depending on the presence of TM9 homologues from both yeast and Arabidopsis thaliana. A yeast tmn1-3 triple mutant lacking all three yeast endogenous TMNs showed altered metal homeostasis with a reduction in the cellular Cu contents to 25% of that in the wild-type. Conversely, when TMN1 was overexpressed in yeast, cellular Cu concentrations were more than doubled. Both Tmn1p-GFP and Tmn2p-GFP fusion proteins localized to the tonoplast. Yeast vacuolar biogenesis was not affected by the lack or presence of TM9 proteins neither in the tmn1-3 triple mutant nor in TM9 overexpressing strains. Heterologous expression in yeast of AtTMN7, a TM9 homologue from Arabidopsis, affected Cu homeostasis similar to the overexpression of TMN1. In Arabidopsis, the two TM9 homologues AtTMN1 and AtTMN7 were ubiquitously expressed. AtTMN7 promoter constructs driving the expression of GFP showed elevated expression of AtTMN7 in the root elongation zone. It is concluded that TM9 homologues from S. cerevisiae and A. thaliana have the ability to affect the intracellular Cu balance. Tmn1p and Tmn2p operate from the yeast vacuolar membrane without influencing vacuolar biogenesis. A new physiological function of the TM9 family coupled to vacuolar Cu homeostasis is proposed.

KW - BRIC

U2 - 10.1111/j.1399-3054.2010.01404.x

DO - 10.1111/j.1399-3054.2010.01404.x

M3 - Journal article

C2 - 20681974

SN - 0031-9317

VL - 140

SP - 355

EP - 367

JO - Physiologia Plantarum

JF - Physiologia Plantarum

IS - 4

ER -