Document Detail

Underexpression of the 43 kDa inositol polyphosphate 5-phosphatase is associated with cellular transformation.
MedLine Citation:
PMID:  8890159     Owner:  NLM     Status:  MEDLINE    
The 43 kDa inositol polyphosphate 5-phosphatase (5-phosphatase) hydrolyses the second messenger molecules inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] and inositol 1,3,4,5-tetrakisphosphate [Ins(1,3,4,5)P4]. We have underexpressed the 43 kDa 5-phosphatase by stably transfecting normal rat kidney cells with the cDNA encoding the enzyme, cloned in the antisense orientation into the tetracycline-inducible expression vector pUHD10-3. Antisense-transfected cells demonstrated a 45% reduction in Ins(1,4,5)P3 5-phosphatase activity in the total cell homogenate upon withdrawal of tetracycline, and an approximately 80% reduction in the detergent-soluble membrane fraction of the cell, as compared with antisense-transfected cells in the presence of tetracycline. Unstimulated antisense-transfected cells showed a concomitant 2-fold increase in Ins(1,4,5)P3 and 4-fold increase in Ins(1,3,4,5)P4 levels. The basal intracellular calcium concentration of antisense-transfected cells (170 +/- 25 nM) was increased 1.9-fold, compared with cells transfected with vector alone (90 +/- 25 nM). Cells underexpressing the 43 kDa 5-phosphatase demonstrated a transformed phenotype. Antisense-transfected cells grew at a 1.7-fold faster rate, reached confluence at higher density and demonstrated increased [3H]thymidine incorporation compared with cells transfected with vector alone. Furthermore, antisense-transfected cells formed colonies in soft agar and tumours in nude mice. These studies support the contention that a decrease in Ins(1,4,5)P3 5-phosphatase activity is associated with cellular transformation.
C J Speed; P J Little; J A Hayman; C A Mitchell
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The EMBO journal     Volume:  15     ISSN:  0261-4189     ISO Abbreviation:  EMBO J.     Publication Date:  1996 Sep 
Date Detail:
Created Date:  1996-12-10     Completed Date:  1996-12-10     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  8208664     Medline TA:  EMBO J     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  4852-61     Citation Subset:  IM    
Department of Medicine, Monash Medical School, Box Hill Hospital, Melbourne, Australia.
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MeSH Terms
3T3 Cells
Calcium / metabolism
Cell Transformation, Neoplastic / metabolism*
Chromatography, High Pressure Liquid
DNA, Complementary / metabolism
Fibroblasts / metabolism
Genes, Tumor Suppressor
Kidney / metabolism
Mice, Nude
Oligonucleotides, Antisense / metabolism
Phosphoric Monoester Hydrolases / biosynthesis*,  genetics
Plasmids / metabolism
Reg. No./Substance:
0/DNA, Complementary; 0/Oligonucleotides, Antisense; 7440-70-2/Calcium; EC 3.1.3.-/Phosphoric Monoester Hydrolases; EC,4,5-trisphosphate 5-phosphatase

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