Developmental regulation of the translational repressor NAT1 during cardiac development.
The process of translation initiation has been postulated to play an important role in the regulation of cellular growth and proliferation. Here, we report the identification and differential expression of a fundamental translational repressor NAT1, during early postnatal cardiac development. Differential display analysis of RNA obtained from 3-day and 4-week-old rat hearts resulted in the cloning and identification of a 396 bp cDNA fragment (DRCF-6) which corresponded to the 3' terminal portion of NAT1. Northern blot analysis revealed that the mRNA expression of NAT1 was markedly elevated during the first 2 weeks of postnatal life, with an apparent peak level of expression occurring at 1 week. NAT1 mRNA levels then steadily decreased to 4 weeks of age. The NAT1 transcript has previously been shown to be extensively edited by the enzyme APOBEC-1, which deaminates specific cytidine bases to uridine; cytidine deamination at a glutamine codon (CAA) results in the formation of a stop codon (UAA) and consequently, premature termination of translation. Accordingly, Western blot analysis detected the presence of several smaller proteins in addition to the full length NAT1 protein (97 kDa), each exhibiting a distinct pattern of expression during cardiac development. APOBEC-1 editing of NAT1 during cardiac development was further supported by primer extension analysis of cytidine 1699, which was found to be predominantly edited to uridine. Immunohistochemical staining showed that NAT1 is expressed predominantly in atrial and ventricular myocytes, although staining was also detected in vascular smooth muscle cells and in the endocardium. These results suggest that NAT1 may play a role in the postnatal development of the heart and demonstrate that APOBEC-1 editing may possibly be a novel mechanism by which translation is regulated during cardiac development.
Pak
B J
BJ
Department of Anatomy and Cell Biology, Queen's University, Kingston, Ontario, K7L 3N6, Canada.
Pang
S C
SC
D000375:Aging; D000818:Animals; D001191:Arylamine N-Acetyltransferase / Q000032:analysis / Q000235:genetics*; D001483:Base Sequence; D018076:DNA, Complementary; D005260:Female; D018507:Gene Expression Regulation, Developmental; D006321:Heart / Q000254:growth & development*; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000032:analysis / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D024510:Muscle Development; D009131:Muscle, Smooth, Vascular / Q000201:enzymology / Q000254:growth & development; D009206:Myocardium / Q000166:cytology / Q000201:enzymology*; D011247:Pregnancy; D051381:Rats; D017208:Rats, Wistar
D018076:DNA, Complementary; D007527:Isoenzymes; D001191:Arylamine N-Acetyltransferase; C089384:N-acetyltransferase 1