NucleaRDB: Extraction of mutation data from the literature

This data was extracted from Medline abstracts and full texts (when available) in an automated manner.

The table below describes the selected point mutation and provides links to other documents. The sentence(s) where the point mutation S112A was found are listed after the table.

The mutated residues are also indicated in the family sequence alignments and hyperlinked to the corresponding mutation pages.

Point mutation S112A in PPAT_HUMAN

Relevant sentences:

S112A

• Adenovirus-mediated gene transfer of either wild-type human peroxisome proliferator-activated receptor (PPAR)-{ gamma}2 or a mutant carrying a replacement at the consensus mitogen-activated protein kinase phosphorylation site (hPPAR-{ gamma}2-S112A) promoted adipogenesis of 3T3-L1 fibroblasts and restored TNF-{ alpha}-induced decrease of triglyceride in adipocytes as effectively as pioglitazone

• The S112A point mutation was introduced into the full-length human PPAR-{ gamma}2 cDNA using QuikChange site-directed mutagenesis kit (Stratagene , La Jolla , CA)

• Briefly , the full-length human PPAR-{ gamma}2 cDNA (either wild-type or S112A mutant) was excised by digestion with SmaI and was inserted into the unique SwaI site of the full-length adenovirus genome cloned in a cassette cosmid , pAxCAwt

• Therefore , to evaluate the role of PPAR-{ gamma} and its adipogenic activity in the insulin-sensitizing effects of TZDs against TNF-{ alpha}-induced insulin resistance , we examined the effects of overexpression of wild-type hPPAR-{ gamma}2 as well as hPPAR-{ gamma}2-S112A , which carries a replacement of the serine to alanine at the consensus MAP kinase phosphorylation site and is assumed to be resistant to phosphorylation by MAP kinases

• Overexpression of S112A mutant hPPAR-{ gamma}2 converted more cells (~60%) into adipocytes (Fig. 5A ) and accumulated more TG in the cells (Fig. 5B )

• These results are similar to those described for 3T3-L1 cells stably expressing mPPAR-{ gamma}2 or mPPAR-{ gamma}2-S112A using a retroviral expression strategy (30 ) , and they indicate that the adenovirus constructs produce PPAR-{ gamma} molecules with expected activities to induce adipocyte differentiation

• Confluent 3T3-L1 fibroblasts were infected with adenovirus vectors containing ß-galactosidase (LacZ) , hPPAR-{ gamma}2 , or hPPAR-{ gamma}2-S112A cDNA at a multiplicity of infection of 50

• Overexpression of either wild-type hPPAR-{ gamma}2 or hPPAR-{ gamma}2-S112A also increased the TG content in the TNF-{ alpha}-treated adipocytes as effectively as pioglitazone (Fig. 6 )

• The cells were infected with adenoviruses containing wild-type (WT) hPPAR-{ gamma}2 , hPPAR-{ gamma}2-S112A , or control ß-galactosidase (LacZ) cDNA at a multiplicity of infection of 50 , 48 h before the measurement of TG content in the adipocytes as described under RESEARCH DESIGN AND METHODS

• Infection of the cells with wild-type or S112A mutant hPPAR-{ gamma}2 adenovirus vector resulted in increases in PPAR-{ gamma}2 levels approximately threefold of the control levels (Fig. 7A )

• Overexpression of either wild-type or S112A mutant hPPAR-{ gamma}2 restored the amounts of IR and IRS-1 protein to the levels comparable with those in control cells (Fig. 7 )

• In contrast , the decreases in insulin-stimulated tyrosine phosphorylation of IR and IRS-1 in TNF-{ alpha}-treated cells were not restored by overexpression of wild-type hPPAR-{ gamma}2 or hPPAR-{ gamma}2-S112A (Fig. 7 )

• The cells were infected with adenoviruses containing wild-type (WT) hPPAR-{ gamma}2 , hPPAR-{ gamma}2-S112A , or control ß-galactosidase (LacZ) cDNA at a multiplicity of infection of 50 , 48 h before the stimulation with insulin (20 nmol / l) for 5 min

• The cells were infected with adenoviruses containing wild-type (WT) hPPAR-{ gamma}2 , hPPAR-{ gamma}2-S112A , or control ß-galactosidase (LacZ) cDNA at a multiplicity of infection of 50 , 48 h before the stimulation with insulin (20 nmol / l) for 15 min

• However , such a mechanism is unlikely to account for the failure to restore tyrosine phosphorylation of IR / IRS-1 , because overexpression of hPPAR-{ gamma}2-S112A , which is assumed to be resistant to serine phosphorylation and has more potent adipogenic activity than wild-type hPPAR-{ gamma}2 , also did not restore tyrosine phosphorylation of IR / IRS-1 and 2-DOG uptake

• Adenovirus-mediated gene transfer of either wild-type human peroxisome proliferator­activated receptor (PPAR)- 2 or a mutant carrying a replacement at the consensus mitogen-activated protein kinase phosphorylation site (hPPAR- 2-S112A) promoted adipogenesis of 3T3-L1 fibroblasts and restored TNF- ­induced decrease of triglyceride in adipocytes as effectively as pioglitazone

• The S112A point mutation was introduced into the full-length human PPAR- 2 cDNA using QuikChange site-directed mutagenesis kit (Stratagene , La Jolla , CA)

• Briefly , the full-length human PPAR- 2 cDNA (either wild-type or S112A mutant) was excised by digestion with SmaI and was inserted into the unique SwaI site of the full-length adenovirus genome cloned in a cassette cosmid , pAxCAwt

• Therefore , to evaluate the role of PPAR- and its adipogenic activity in the insulin-sensitizing effects of TZDs against TNF- ­induced insulin resistance , we examined the effects of overexpression of wildtype hPPAR- 2 as well as hPPAR- 2-S112A , which carries a replacement of the serine to alanine at the consensus MAP kinase phosphorylation site and is assumed to be resistant to phosphorylation by MAP kinases

• Overexpression of S112A mutant hPPAR- 2 converted more cells ( 60%) into adipocytes (Fig. 5A) and accumulated more TG in the cells (Fig. 5B)

• These results are similar to those described for 3T3-L1 cells stably expressing mPPAR- 2 or mPPAR- 2-S112A using a retroviral expression strategy (30) , and they indicate that the adenovirus constructs produce PPAR- molecules with expected activities to induce adipocyte differentiation

• Overexpression of either wild-type hPPAR- 2 or hPPAR- 2-S112A also increased the TG content in the TNF- ­treated adipocytes as effectively as pioglitazone (Fig. 6)

• Infection of the cells with wild-type or S112A mutant hPPAR- 2 adenovirus vector resulted in increases in PPAR- 2 levels approximately threefold of the control levels (Fig. 7A)

• Overexpression of either wild-type or S112A mutant hPPAR- 2 restored the amounts of IR and IRS-1 protein to the levels comparable with those in control cells (Fig. 7)

• In contrast , the decreases in insulinstimulated tyrosine phosphorylation of IR and IRS-1 in TNF- ­treated cells were not restored by overexpression of wild-type hPPAR- 2 or hPPAR- 2-S112A (Fig. 7)

• Confluent 3T3-L1 fibroblasts were infected with adenovirus vectors containing -galactosidase (LacZ) , hPPAR- 2 , or hPPAR- 2-S112A cDNA at a multiplicity of infection of 50

• However , such a mechanism is unlikely to account for the failure to restore tyrosine phosphorylation of IR / IRS-1 , because overexpression of hPPAR- 2-S112A , which is assumed to be resistant to serine phosphorylation and has more potent adipogenic activity than wild-type hPPAR- 2 , also did not restore tyrosine phosphorylation of IR / IRS-1 and 2-DOG uptake

• The cells were infected with adenoviruses containing wild-type (WT) hPPAR- 2 , hPPAR- 2-S112A , or control -galactosidase (LacZ) cDNA at a multiplicity of infection of 50 , 48 h before the measurement of TG content in the adipocytes as described under RESEARCH DESIGN AND METHODS

• The cells were infected with adenoviruses containing wild-type (WT) hPPAR- 2 , hPPAR- 2-S112A , or control -galactosidase (LacZ) cDNA at a multiplicity of infection of 50 , 48 h before the stimulation with insulin (20 nmol / l) for 5 min

F.Horn (nucleardbcmbi.kun.nl), 21-Apr-2005