Functional significance of human trp1 and trp3 in store-operated Ca2+ entry in HEK-293 cells

被引:98
作者
Wu, XY [1 ]
Babnigg, G [1 ]
Villereal, ML [1 ]
机构
[1] Univ Chicago, Dept Neurobiol Pharmacol & Physiol, Chicago, IL 60637 USA
来源
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY | 2000年 / 278卷 / 03期
关键词
human transient receptor potential proteins; transient receptor potential; antisense cDNA; calcium store depletion; thapsigargin; Ba2+ influx; stable transfection; human embryonic kidney cells;
D O I
10.1152/ajpcell.2000.278.3.C526
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The Drosophila trp (transient receptor potential) gene appears to encode the Drosophila store-operated channel(SOC), and some mammalian trp homologues have been proposed to encode mammalian SOCs. This study provides evidence for the expression of three trp homologues (Mtrp2, Mtrp3, and Mtrp4) in fibroblasts from wild-type and src knockout mice, and four trp homologues (Htrp1, Htrp3, Htrp4, and Htrp6) in human embryonic kidney (HEK-293) cells based on RT-PCR techniques. In HEK-293 cells stably transfected with a 323-bp Htrp3 antisense construct (Htrp3AS), Northern blot analysis revealed that the expression of a 4-kb transcript was dramatically suppressed in comparison to that observed in cells stably transfected with a short Htrp3 sense construct (Htrp3S). Activity of SOCs, monitored as Ba2+ influx following Ca2+ store depletion with thapsigargin, was reduced by 32% in Htrp3AS cells in comparison with Htrp3S cells. Transient transfection of a 369-bp Htrp1 antisense construct in cells stably expressing Htrp3AS induced a higher level of inhibition (55%) of store-operated Ca2+ entry. These data suggest that Htrp1 and Htrp3 may be functional subunits of SOCs.
引用
收藏
页码:C526 / C536
页数:11
相关论文
共 58 条
[31]   Store-operated calcium entry promotes shape change in pulmonary endothelial cells expressing Trp1 [J].
Moore, TM ;
Brough, GH ;
Babal, P ;
Kelly, JJ ;
Li, M ;
Stevens, T .
AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, 1998, 275 (03) :L574-L582
[32]   Molecular cloning of a novel putative Ca2+ channel protein (TRPC7) highly expressed in brain [J].
Nagamine, K ;
Kudoh, J ;
Minoshima, S ;
Kawasaki, K ;
Asakawa, S ;
Ito, F ;
Shimizu, N .
GENOMICS, 1998, 54 (01) :124-131
[33]  
NUOVO G, 1995, PRIMER LAB MANUAL
[34]   Molecular cloning and functional characterization of a novel receptor-activated TRP Ca2+ channel from mouse brain [J].
Okada, T ;
Shimizu, S ;
Wakamori, M ;
Maeda, A ;
Kurosaki, T ;
Takada, N ;
Imoto, K ;
Mori, Y .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (17) :10279-10287
[35]   Store depletion and calcium influx [J].
Parekh, AB ;
Penner, R .
PHYSIOLOGICAL REVIEWS, 1997, 77 (04) :901-930
[36]   REGULATION OF CALCIUM INFLUX BY 2ND MESSENGERS IN RAT MAST-CELLS [J].
PENNER, R ;
MATTHEWS, G ;
NEHER, E .
NATURE, 1988, 334 (6182) :499-504
[37]   PUTATIVE CAPACITATIVE CALCIUM-ENTRY CHANNELS - EXPRESSION OF DROSOPHILA TRP AND EVIDENCE FOR THE EXISTENCE OF VERTEBRATE HOMOLOGS [J].
PETERSEN, CCH ;
BERRIDGE, MJ ;
BORGESE, MF ;
BENNETT, DL .
BIOCHEMICAL JOURNAL, 1995, 311 :41-44
[38]   A mammalian capacitative calcium entry channel homologous to Drosophila TRP and TRPL [J].
Philipp, S ;
Cavalie, A ;
Freichel, M ;
Wissenbach, U ;
Zimmer, S ;
Trost, C ;
Marquart, A ;
Murakami, M ;
Flockerzi, V .
EMBO JOURNAL, 1996, 15 (22) :6166-6171
[39]   A novel capacitative calcium entry channel expressed in excitable cells [J].
Philipp, S ;
Hambrecht, J ;
Braslavski, L ;
Schroth, G ;
Freichel, M ;
Murakami, M ;
Cavalié, A ;
Flockerzi, V .
EMBO JOURNAL, 1998, 17 (15) :4274-4282
[40]   Expression and characterization of a trpl homolog from rat [J].
Preuss, KD ;
Noller, JK ;
Krause, E ;
Gobel, A ;
Schulz, I .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1997, 240 (01) :167-172