title: A porphobilinogen deaminase (PBGD) Ran-binding protein interaction is implicated in nuclear trafficking of PBGD in differentiating glioma cells.
abstract: Porphobilinogen deaminase (PBGD) is a rate-limiting enzyme of the heme biosynthesis pathway, whose level is elevated in various human tumors. PBGD was observed in both nuclear and cytoplasmic fractions of C6 glioma cells by immunostaining. During mitosis, chromatids were intensely stained for PBGD in comparison to the interphase chromatin. Using the yeast two-hybrid system, we identified RanBPM, the nuclear Ran-binding protein, as an interacting partner of PBGD. During butyrate-induced differentiation of C6, both nuclear and cytoplasmic PBGD levels declined as did Ran protein and its nucleotide exchange factor RCC1. N,N'-hexamethylene bis-acetamide-dependent differentiation resulted in an increase of the cytoplasmic PBGD, whereas nuclear PBGD, Ran protein and RCC1 remained unchanged. mRNA levels of PBGD remained unchanged during stimulation with both butyrate and N,N'-hexamethylene bis-acetamide. The enzymatic activity of PBGD and protoporphyrin IX synthesis in C6 cells were dependent on the differentiation induction agent. We conclude that PBGD possibly has a nuclear role in addition to its cytosolic enzymatic activity required for heme synthesis, which is related to cell transformation and differentiation.
authors.affiliation: Life Science Faculty, Bar-Ilan University, Ramat-Gan 52900, Israel.
authors.forename: Lior
authors.initials: L
authors.lastname: Greenbaum
authors.forename: Don J
authors.initials: DJ
authors.lastname: Katcoff
authors.forename: Huanyu
authors.initials: H
authors.lastname: Dou
authors.forename: Yael
authors.initials: Y
authors.lastname: Gozlan
authors.forename: Zvi
authors.initials: Z
authors.lastname: Malik
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002467:Cell Nucleus / Q000378:metabolism*; D005910:Glioma / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D011163:Hydroxymethylbilane Synthase / Q000378:metabolism*; D008969:Molecular Sequence Data; D011166:Porphyrins / Q000378:metabolism; D051381:Rats; D020931:ran GTP-Binding Protein / Q000378:metabolism*
chemical_list: D011166:Porphyrins; D011163:Hydroxymethylbilane Synthase; D020931:ran GTP-Binding Protein