title: Tissue-dependent loss of phosphofructokinase-M in mice with interrupted activity of the distal promoter: impairment in insulin secretion.
abstract: Phosphofructokinase is a key enzyme of glycolysis that exists as homo- and heterotetramers of three subunit isoforms: muscle, liver, and C type. Mice with a disrupting tag inserted near the distal promoter of the phosphofructokinase-M gene showed tissue-dependent differences in loss of that isoform: 99% in brain and 95-98% in islets, but only 50-75% in skeletal muscle and little if any loss in heart. This correlated with the continued presence of proximal transcripts specifically in muscle tissues. These data strongly support the proposed two-promoter system of the gene, with ubiquitous use of the distal promoter and additional use of the proximal promoter selectively in muscle. Interestingly, the mice were glucose intolerant and had somewhat elevated fasting and fed blood glucose levels; however, they did not have an abnormal insulin tolerance test, consistent with the less pronounced loss of phosphofructokinase-M in muscle. Isolated perifused islets showed about 50% decreased glucose-stimulated insulin secretion and reduced amplitude and regularity of secretory oscillations. Oscillations in cytoplasmic free Ca(2+) and the rise in the ATP/ADP ratio appeared normal. Secretory oscillations still occurred in the presence of diazoxide and high KCl, indicating an oscillation mechanism not requiring dynamic Ca(2+) changes. The results suggest the importance of phosphofructokinase-M for insulin secretion, although glucokinase is the overall rate-limiting glucose sensor. Whether the Ca(2+) oscillations and residual insulin oscillations in this mouse model are due to the residual 2-5% phosphofructokinase-M or to other phosphofructokinase isoforms present in islets or involve another metabolic oscillator remains to be determined.
authors.affiliation: Department of Biochemistry, Boston University School of Medicine, Boston, MA 02118, USA.
authors.forename: Ann-Marie T
authors.initials: AM
authors.lastname: Richard
authors.forename: Dominic-Luc
authors.initials: DL
authors.lastname: Webb
authors.forename: Jessie M
authors.initials: JM
authors.lastname: Goodman
authors.forename: Vera
authors.initials: V
authors.lastname: Schultz
authors.forename: John N
authors.initials: JN
authors.lastname: Flanagan
authors.forename: Lisa
authors.initials: L
authors.lastname: Getty-Kaushik
authors.forename: Jude T
authors.initials: JT
authors.lastname: Deeney
authors.forename: Gordon C
authors.initials: GC
authors.lastname: Yaney
authors.forename: George A
authors.initials: GA
authors.lastname: Dunaway
authors.forename: Per-Olof
authors.initials: PO
authors.lastname: Berggren
authors.forename: Keith
authors.initials: K
authors.lastname: Tornheim
mesh_terms: D000818:Animals; D001786:Blood Glucose / Q000378:metabolism*; D006014:Glycogen Storage Disease Type VII / Q000235:genetics* / Q000378:metabolism*; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D008657:Metabolic Clearance Rate; D051379:Mice; D008822:Mice, Transgenic; D009928:Organ Specificity; D010732:Phosphofructokinase-1 / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014018:Tissue Distribution
chemical_list: D001786:Blood Glucose; D007328:Insulin; D010732:Phosphofructokinase-1