Characterization of the Core Promoter of the Na+/K+‐ATPase α1 Subunit Gene

FEBS Journal - Tập 237 Số 2 - Trang 440-446 - 1996
Kiyoshi Kawakami1, Kazuyuki Masuda1, Kenichi Nagano1, Yoshiaki Ohkuma2, Robert G. Roeder2
1Department of Biology, Jichi Medical School, Minamikawachi-machi, Kawachi-gun, Tochigi-ken, Japan
2Laboratory of Biochemistry and Molecular Biology, the Rockefeller University, New York, USA

Tóm tắt

We have analyzed the core promoter element of the Na+/K+‐ATPase α1 subunit gene by means of an in vitro transcription system composed of a HeLa nuclear extract. 5′‐deletion and 3′‐deletion analyses revealed that this gene is specifically transcribed by RNA polymerase II in a manner that is dependent on the upstream regulatory region of the gene (−102 to −61), and that the 3′ boundary of the minimal promoter element does not extend beyond +5. Analysis of linker‐substitution mutations and point mutations revealed that the TATA‐like sequence (−33 to −26) is required for upstream‐sequence‐dependent transcription whereas linker‐substitution mutations and point mutations near +1 did not abolish transcription.The gene was found to be transcribed by RNA polymerase III when phosphocellulose column fractions were assayed. Deletion analysis mapped the minimal RNA‐polymerase‐III–specific promoter element from −49 to +17. The phosphocellulose 0.3‐M‐KCl fraction is absolutely required for transcription by RNA polymerase III, while the 0.85‐M‐KCl fraction represses aberrant transcription from incorrect initiation sites. Analysis of linker‐substitution mutations indicated that the TATA‐like sequence is required for RNA‐polymerase‐III–specific transcription. Although point mutations in the 5′ half of the TATA‐like sequence did not affect transcription, those in the 3′ half shifted the transcription initiation site 3 bp upstream. The results suggest that the Na+/K+‐ATPase α1 subunit gene promoter contains a TATA‐like sequence which can direct transcription by RNA polymerase III in vitro. The mechanism of alternative regulation of RNA polymerase II and RNA polymerase III is discussed.

Từ khóa


Tài liệu tham khảo

Smith T. J., 1979, The role of sodium transport in thyroid thermogenesis, Fed. Proc., 38, 2150

Lingrel J. B., 1992, Na+/K+‐ATPase: isoform structure, function, and expression, J. Bioenerg. Biomembr., 24, 263, 10.1007/BF00768847

Orlowski J., 1988, Tissue‐specific and developmental regulation of rat Na, K‐ATPase catalytic α isoform and β subunit mRNAs, J. Biol. Chem., 263, 10436, 10.1016/S0021-9258(19)81535-1

10.1128/MCB.12.9.4046

10.1093/oxfordjournals.jbchem.a124267

Kawakami K., 1993, Different factors bind to the regulatory region of the Na+ K+‐ATPase α1‐subunit gene during the cell cycle, FEBS Lett., 535, 251, 10.1016/0014-5793(93)80740-L

10.1093/nar/23.15.2848

Kawakami K., 1994, The sodium pump: structure, mechanism, hormonal control and its role in disease, 1

10.1093/nar/11.5.1475

Smale S. T., 1994, Transcription; mechanisms and regulation, 63

Steinberg T. H., 1990, Tagetoxin: a new inhibitor of eukaryotic transcription by RNA polymerase III, J. Biol. Chem., 265, 499, 10.1016/S0021-9258(19)40259-7

Steinberg T. H., 1992, Tagetoxin inhibition of RNA polymerase III transcription results from enhanced pausing at discrete sites and is template‐dependent, J. Biol. Chem., 267, 20204, 10.1016/S0021-9258(19)88687-8

10.1016/0092-8674(87)90012-2

10.1016/0092-8674(89)90402-9

10.1128/MCB.14.1.116

Kassavetis G. A., 1994, Transcription; mechanisms and regulation, 107

Goomer R. S., 1992, The transcriptional start site for a human U6 small nuclear RNA gene is dictated by a compound promoter element consisting of the PSE and the TATA box, Nucleic Acids Res., 18, 4903, 10.1093/nar/20.18.4903

10.1016/0092-8674(89)90763-0

10.1101/gad.3.8.1179

Pruzan R., 1992, Specific transcription from the adenovirus E2E promoter by RNA polymerase III requires a subpopulation of TFIID, Nucleic Acids Res., 20, 5705, 10.1093/nar/20.21.5705

Mitchell M. T., 1992, TATA box‐mediated polymerase III transcription in vitro, J. Biol. Chem., 267, 1995, 10.1016/S0021-9258(18)46045-0

Segall J., 1980, Multiple factors are required for the accurate transcription of purified genes by RNA polymerase III, J. Biol. Chem., 255, 11986, 10.1016/S0021-9258(19)70231-2

10.1101/gad.5.8.1477

Thông tin
: []
Thông tin xuất bản

FEBS Journal

Tập 237 Số 2

440-446

Thông tin tác giả