Cloning, expression and chromosomal localization of a human testis 6- phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene

A. Manzano, J. X. Pérez, M. Nadal, X. Estivill, A. Lange, R. Bartrons

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Abstract

6-Phosphofructo-2-kinase/fructose 2,6-bisphosphatase (PFK-2/FBPase-2) is a bifunctional enzyme responsible for the synthesis and breakdown of Fru- 2,6-P2, a key metabolite in the regulation of glycolysis. Several genes encode distinct PFK-2/FBPase-2 isozymes that differ in their tissue distribution and enzyme regulation. In this paper, we present the isolation of a cDNA from a human testis cDNA library that encodes a PFK-2/FBPase-2 isozyme. Sequencing data show an open reading frame of 1407 nucleotides that codifies for a protein of 469 amino acids. This has a calculated molecular weight of 54 kDa and 97% similarity with rat testis PFK-2/FBPase-2, with complete conservation of the amino acid residues involved in the catalytic mechanism. Fluorescence in-situ hybridization (FISH) localized testis PFK- 2/FBPase-2 gene (PFKFB4) in human chromosome 3 at bands p21-p22. A Northern blot analysis of different rat tissues showed the presence of a 2.4-kb mRNA expressed specifically in testis. In mammalian COS-1 cells, the human testis cDNA drives expression of an isozyme with a molecular weight of 55 kDa. This isozyme shows clear PFK-2 activity. Taken together, these results provide evidence for a new PFK-2/FBPase-2 gene coding for a human testis isozyme.

Original languageEnglish (US)
Pages (from-to)83-89
Number of pages7
JournalGene
Volume229
Issue number1-2
DOIs
StatePublished - Mar 18 1999

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Keywords

  • 6-phosphofructo-2-kinase/fructose 2,6- bisphosphatase
  • Gene
  • Glycolysis
  • Testis

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