Isolation and Cloning of Rat Poly (ADP-Ribose) Glycohydrolase: Presence of a Potential Nuclear Export Signal Conserved in Mammalian Orthologs

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Author(s)

Abstract

Poly (ADP-ribose) glycohydrolase (Parg) is the main enzyme of poly (ADP-ribose) degradation. To understand its structure-and-function relationship, we purified Parg from rat testis 9, 740-fold using an improved affinity column; the purified product was a 60 kDa protein. Based on the determined sequences of three peptide fragments, degenerated primers were synthesized and a <i>Parg</i> cDNA comprising 3, 974 nucleotides, encoding a 109 kDa protein, was isolated. The 60 kDa Parg purified from rat testes corresponded to the C-terminal half of the 109 kDa deduced peptide. When recombinant rat Parg was expressed as a glutathione S-transferase fusion protein in <i>Escherichia coli</i>, Parg activity was observed for the full-length and C-terminal half proteins but not in for the N-terminal half protein. Taken together, these data indicate that the catalytic domain of Parg is located in the C-terminal half. Further, we newly identified the presence of a potential nuclear export signal in the N-terminal half in addition to the previously reported nuclear localization signals in rat and other mammalian Pargs. Northern blot analysis showed the ubiquitous expression of a single 4.0 kb <i>Parg</i> mRNA in various rat tissues. The findings suggest that the 60 kDa Parg is produced by post-transcriptional processing.

Journal

  • The Journal of Biochemistry

    The Journal of Biochemistry 126(4), 748-755, 1999-10-01

    The Japanese Biochemical Society

References:  38

Codes

  • NII Article ID (NAID)
    10005858785
  • NII NACSIS-CAT ID (NCID)
    AA00694073
  • Text Lang
    ENG
  • Article Type
    ART
  • ISSN
    0021924X
  • NDL Article ID
    4861216
  • NDL Source Classification
    ZR2(科学技術--生物学--生化学)
  • NDL Call No.
    Z53-B472
  • Data Source
    CJP  NDL  J-STAGE 
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