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Endocrinology Vol. 139, No. 4 1999-2006
Copyright © 1998 by The Endocrine Society

ARTICLES

Multiple Transcripts Encoded by the Thyroid-Specific Enhancer-Binding Protein (T/EBP)/Thyroid-Specific Transcription Factor-1 (TTF-1) Gene: Evidence of Autoregulation1

Hidenori Oguchi2 and Shioko Kimura

Laboratory of Metabolism, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892

Address all correspondence and requests for reprints to: Shioko Kimura, Ph.D., Building 37, Room 3E-24, National Institutes of Health, 9000 Rockville Pike, Bethesda, Maryland 20892. E-mail: shioko{at}helix.nih.gov

Multiple transcripts derived from the gene encoding rat thyroid-specific enhancer-binding protein (T/EBP)/thyroid-specific transcription factor-1 (TTIF-1) were identified by complementary DNA cloning and sequencing, and Northern blotting analyses. Six different types of complementary DNAs were identified that differ at their 5' noncoding regions; four contain an intron of different lengths, whereas the other two possess no intron. Ribonuclease protection analyses revealed that multiple promoters are scattered throughout the upstream region, and the usage of these different promoters together with alternative splicing leads to a family of T/EBP messenger RNA (mRNA) species. A similar pattern of expression was also found in the human T/EBP gene expressed in a lung carcinoma cell line. Longer T/EBP mRNAs are more abundant in rat FRTL-5 thyroid cells maintained in the absence of TSH (-TSH) than in cells maintained in the presence of TSH (+TSH). Transfection analyses using the rat T/EBP gene DNA upstream of the ATG initiation codon connected to the luciferase reporter plasmid showed a similar relative activity profile between -TSH and +TSH culture conditions, suggesting that the abundance of longer mRNAs in -TSH conditions may not directly correlate with differences in promoter activities. Rather, TSH status might have a role in maintaining the physiological state of the cells. The upstream DNA of the rat and human T/EBP genes share a cluster of high and low sequence similarities, and both possess respectively 24 and 18 putative T/EBP-binding sites throughout. Cotransfection analyses of the T/EBP promoter-reporter constructs with a T/EBP expression vector into human HepG2 cells, which do not express T/EBP, suggested that autoregulation may be involved in controlling both rat and human T/EBP gene expression.




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