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, Xavier P. Estivill, A. Lange, R. Bartrons

Research output: Contribution to journalArticle

28 Citations (Scopus)

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
Pages (from-to)83-89
Number of pages7
JournalGene
Volume229
Issue number1-2
DOIs
Publication statusPublished - 18 Mar 1999
Externally publishedYes

Fingerprint

Testis
Organism Cloning
Isoenzymes
Genes
Complementary DNA
Molecular Weight
Phosphofructokinase-2
Amino Acids
Chromosomes, Human, Pair 3
COS Cells
Human Chromosomes
Glycolysis
Tissue Distribution
Enzymes
Fluorescence In Situ Hybridization
Gene Library
Northern Blotting
Open Reading Frames
human PFKFB3 protein
Nucleotides

Keywords

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

ASJC Scopus subject areas

  • Genetics

Cite this

Cloning, expression and chromosomal localization of a human testis 6- phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene. / Manzano, A.; Pérez, J. X.; Nadal, M.; Estivill, Xavier P.; Lange, A.; Bartrons, R.

In: Gene, Vol. 229, No. 1-2, 18.03.1999, p. 83-89.

Research output: Contribution to journalArticle

Manzano, A. ; Pérez, J. X. ; Nadal, M. ; Estivill, Xavier P. ; Lange, A. ; Bartrons, R. / Cloning, expression and chromosomal localization of a human testis 6- phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene. In: Gene. 1999 ; Vol. 229, No. 1-2. pp. 83-89.
@article{9b716feeb1c04abcab75c907f299258c,
title = "Cloning, expression and chromosomal localization of a human testis 6- phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene",
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.",
keywords = "6-phosphofructo-2-kinase/fructose 2,6- bisphosphatase, Gene, Glycolysis, Testis",
author = "A. Manzano and P{\'e}rez, {J. X.} and M. Nadal and Estivill, {Xavier P.} and A. Lange and R. Bartrons",
year = "1999",
month = "3",
day = "18",
doi = "10.1016/S0378-1119(99)00037-2",
language = "English",
volume = "229",
pages = "83--89",
journal = "Gene",
issn = "0378-1119",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - Cloning, expression and chromosomal localization of a human testis 6- phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene

AU - Manzano, A.

AU - Pérez, J. X.

AU - Nadal, M.

AU - Estivill, Xavier P.

AU - Lange, A.

AU - Bartrons, R.

PY - 1999/3/18

Y1 - 1999/3/18

N2 - 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.

AB - 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.

KW - 6-phosphofructo-2-kinase/fructose 2,6- bisphosphatase

KW - Gene

KW - Glycolysis

KW - Testis

UR - http://www.scopus.com/inward/record.url?scp=0344923803&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0344923803&partnerID=8YFLogxK

U2 - 10.1016/S0378-1119(99)00037-2

DO - 10.1016/S0378-1119(99)00037-2

M3 - Article

VL - 229

SP - 83

EP - 89

JO - Gene

JF - Gene

SN - 0378-1119

IS - 1-2

ER -