cDNA cloning of rat major AP endonuclease (APEX nuclease) and analyses of its mRNA expression in rat tissues.
APEX nuclease is a mammalian DNA repair enzyme having apurinic/apyrimidinic (AP) endonuclease, 3'-5'-exonuclease, DNA 3' repair diesterase and DNA 3'-phosphatase activities. It is also a redox factor (Ref-1), stimulating DNA binding activity of AP-1 binding proteins such as Fos and Jun. In the present paper, a cDNA for the enzyme was isolated from a rat brain cDNA library using mouse Apex cDNA as a probe and sequenced. The rat Apex cDNA was 1221 nucleotides (nt) long, with a 951-nt coding region. The amino acid sequence of rat APEX nuclease has 98.4% identity with mouse APEX nuclease. Using the rat Apex cDNA as a probe for Northern blot analysis, the size of rat Apex mRNA was shown to be approximately 1.5 kb. Its expression was compared in 9 rat organs on postnatal days 7 and 28. Although Apex mRNA was expressed ubiquitously, the levels varied significantly, suggesting organ- or tissue-specific expression of the Apex gene. The highest level was observed in the testis, relatively high levels in the thymus, spleen, kidney and brain, and the lowest level in the liver. The level of expression at postnatal day 28, with the exception of the testis, was almost the same as or lower in respective organs than that at postnatal day 7. Postnatal developmental changes of Apex mRNA expression in the testis and thymus were further studied. The expression in testis was markedly increased on postnatal days 21 and 28. The expression in thymus increased once at postnatal day 14, and then decreased. The developmental changes of Apex mRNA expression in testis and thymus suggest that APEX nuclease is involved in processes such as recombinational events.
Tan
Y
Y
Department of Molecular Biology, Institute of Cellular and Molecular Biology, Okayama University Medical School, Japan.
Nakagawa
Y
Y
Akiyama
K
K
Wakabayashi
H
H
Sarker
A H
AH
Seki
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019757:Carbon-Oxygen Lyases; D003001:Cloning, Molecular; D004260:DNA Repair; D018076:DNA, Complementary / Q000302:isolation & purification*; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D004706:Endodeoxyribonucleases / Q000235:genetics*; D005090:Exodeoxyribonucleases / Q000235:genetics*; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics*; D009687:Nuclear Proteins / Q000378:metabolism*; D009928:Organ Specificity; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D011919:Rats, Inbred Strains; D013737:Testis / Q000254:growth & development / Q000378:metabolism; D013950:Thymus Gland / Q000254:growth & development / Q000378:metabolism
D018076:DNA, Complementary; D009097:Multienzyme Complexes; D009687:Nuclear Proteins; D012333:RNA, Messenger; D004706:Endodeoxyribonucleases; D005090:Exodeoxyribonucleases; D019757:Carbon-Oxygen Lyases; C486216:Apex1 protein, mouse; C470319:Apex1 protein, rat; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase