Molecular cloning, expression, and chromosomal localization of a ubiquitously expressed human 6-phosphofructo-2-kinase/ fructose-2,6-bisphosphatase gene (PFKFB3)

Abstract

We report the identification of a human 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase gene (PFKFB3) isolated from a human fetal brain cDNA library. The gene was localized to 10p15→p14 by fluorescence in situ hybridization. The entire cDNA (4,322 bp) codes for a polypeptide of 520 amino acid residues (molecular weight, 59.571 kDa). Structural analysis showed the presence of a kinase domain located at the amino terminus and a bisphosphatase domain at the carboxy terminus, characteristic of previously described 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase isozymes. In addition, a phosphorylation site for cAMP-dependent protein kinase was found at the carboxy terminus. Northern blot analysis showed the presence of a unique 4.8-kb mRNA expressed in the different tissues studied. In mammalian COS-1 cells, this cDNA drives the expression of an active isozyme. Taken together, these results identify the presence of a gene coding for a human 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase isozyme which is ubiquitously expressed.

RNAi screening in glioma stem-like cells identifies PFKFB4 as a key molecule important for cancer cell survival

“…It has been originally identified in the testes and shown to be highly expressed in both colon and breast cancers (Minchenko et al, 2004;Gomez et al, 2005). The homologous PFKFB3 encodes an inducible form of PFK2/FBPase-2 that is ubiquitously expressed in many normal tissues, such as brain and liver, and overexpressed in different cancers (Manzano et al, 1998;Kessler and Eschrich, 2001;Atsumi et al, 2002). Both PFKFB3 and PFKFB4 are induced by hypoxia in various tumors.…”

“…Statistical significance was calculated using Student's t-test for ranked transformed data. Brain cancer stem-like cells require PFKFB4 for survival V Goidts et al another isozyme of PFK2 that is expressed in normal brain and overexpressed in numerous tumors (Manzano et al, 1998;Kessler and Eschrich, 2001;Atsumi et al, 2002;Gomez-Brouchet et al, 2007). As depicted in Figure 3a, PFKFB3 showed a 1.3-fold higher mean mRNA expression level in primary glioblastomas relative to normal brain tissue and was not overexpressed in the other glioma groups investigated.…”

6-Phosphofructo-2-kinase (pfkfb3) Gene Promoter Contains Hypoxia-inducible Factor-1 Binding Sites Necessary for Transactivation in Response to Hypoxia

“…These reagents inhibit HIF-1 prolyl hydroxylases activity (7). Human glioblastoma T98G and U-87 cell lines were chosen because of positive gene expression for pfkfb3 (22). Western blot results from T98G (Fig.…”

Ras transformation requires metabolic control by 6-phosphofructo-2-kinase

“…The enzymes derived from PFKFB1, PFKFB2 and PFKFB4 were originally found to be expressed by cells of the liver/muscle, kidney/heart and testes, respectively, and all display nearly equal kinase: phosphatase ratios (Okar and Lange, 1999). The PFKFB3 gene encodes for the inducible PFK2/FBPase (Chesney et al, 1999;Atsumi et al, 2002;Mahlknecht et al, 2003), which is rapidly induced by inflammatory stimuli (Chesney et al, 1999) and hypoxia (Minchenko et al, 2002;Minchenko et al, 2004) and has also been termed placental PFK2 (Sakai et al, 1996;Sakakibara et al, 1999), ubiquitous PFK2 (Manzano et al, 1998;Kessler and Eschrich, 2001;Navarro-Sabate et al, 2001) and PGR1 (Hamilton et al, 1997)). The PFKFB3 protein product was recently documented to be overexpressed in the neoplastic cells of human solid tumors, including lung, breast, prostate and colon tumors and to display minimal phosphatase activity (kinase:phosphatase ratio 740:1), suggesting a constitutively pro-glycolytic activity (Sakakibara et al, 1997;Chesney et al, 1999;Chesney and Bucala, 2001;Atsumi et al, 2002).…”