» Articles » PMID: 8573111

Differential Transcription of the Human Spermidine/spermine N1-acetyltransferase (SSAT) Gene in Human Lung Carcinoma Cells

Overview
Journal Biochem J
Specialty Biochemistry
Date 1996 Jan 15
PMID 8573111
Citations 12
Authors
Affiliations
Soon will be listed here.
Abstract

The expression of spermidine/spermine N1-acetyltransferase (SSAT), the rate-limiting enzyme in the catabolism of polyamines, is highly regulated by a number of factors including the natural polyamines and their analogues. The phenotype-specific cytotoxicity that occurs in response to a class of polyamine analogues, the diethylpolyamines, is associated with a phenotype-specific superinduction of SSAT in human non-small-cell lung carcinomas, whereas in non-responding cell types, including the small-cell lung carcinomas, the superinduction of SSAT does not occur. In this study, we have investigated the molecular basis of this phenotype-specific SSAT induction in human lung carcinoma cells in response to N1,N12-diethylspermine (BESpm). To facilitate the study of transcriptional regulation, we have cloned and characterized 11 kb of the human SSAT locus, including 3500 bp of the 5' promoter region. Nuclear run-on transcription studies suggest that the initial induction of SSAT results from an increase in the rate of gene transcription. Results from Northern blot analysis and ribonuclease protection assays indicate a differential expression of SSAT mRNA between the analogue-responsive H157 and non-responsive H82 cells. There is no detectable SSAT mRNA in H82 cells, even after a 24-h analogue treatment, whereas SSAT mRNA in H157 cells was detectable by Northern blot analysis and increased more than 100-fold following drug exposure. Furthermore, nuclear run-on transcription assays do not detect any active transcription of SSAT gene in either treated or untreated H82 cells. These results indicate that at least one component of the phenotype-specific induction of SSAT appears to be due to differences in transcriptional regulation of the gene. In addition, mapping of DNase I-hypersensitive sites of the SSAT gene suggest that the cell type-specific promoter/enhancer utilization may control the expression of the SSAT gene in differentially sensitive cell types in vivo.

Citing Articles

Specific phosphorylation of histone demethylase KDM3A determines target gene expression in response to heat shock.

Cheng M, Zhang Y, Cao C, Zhang W, Zhang Y, Shen Y PLoS Biol. 2014; 12(12):e1002026.

PMID: 25535969 PMC: 4275180. DOI: 10.1371/journal.pbio.1002026.


Histone deacetylase inhibition overcomes drug resistance through a miRNA-dependent mechanism.

Murray-Stewart T, Hanigan C, Woster P, Marton L, Casero Jr R Mol Cancer Ther. 2013; 12(10):2088-99.

PMID: 23943804 PMC: 3808125. DOI: 10.1158/1535-7163.MCT-13-0418.


Downregulation of the polyamine regulator spermidine/spermine N(1)-acetyltransferase by Epstein-Barr virus in a Burkitt's lymphoma cell line.

Shi M, Gan Y, Davis T, Scott R Virus Res. 2013; 177(1):11-21.

PMID: 23891576 PMC: 3852694. DOI: 10.1016/j.virusres.2013.07.004.


Heat shock protein 10 and signal transduction: a "capsula eburnea" of carcinogenesis?.

Czarnecka A, Campanella C, Zummo G, Cappello F Cell Stress Chaperones. 2007; 11(4):287-94.

PMID: 17278877 PMC: 1713189. DOI: 10.1379/csc-200.1.


Polyamine-regulated unproductive splicing and translation of spermidine/spermine N1-acetyltransferase.

Hyvonen M, Uimari A, Keinanen T, Heikkinen S, Pellinen R, Wahlfors T RNA. 2006; 12(8):1569-82.

PMID: 16809818 PMC: 1524884. DOI: 10.1261/rna.39806.


References
1.
Feinberg A, Vogelstein B . "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984; 137(1):266-7. DOI: 10.1016/0003-2697(84)90381-6. View

2.
Southern E . Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975; 98(3):503-17. DOI: 10.1016/s0022-2836(75)80083-0. View

3.
Melton D, Krieg P, Rebagliati M, Maniatis T, Zinn K, Green M . Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984; 12(18):7035-56. PMC: 320141. DOI: 10.1093/nar/12.18.7035. View

4.
Porter C, McManis J, Casero R, Bergeron R . Relative abilities of bis(ethyl) derivatives of putrescine, spermidine, and spermine to regulate polyamine biosynthesis and inhibit L1210 leukemia cell growth. Cancer Res. 1987; 47(11):2821-5. View

5.
Chomczynski P, Sacchi N . Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987; 162(1):156-9. DOI: 10.1006/abio.1987.9999. View