Nucleic Acids Research, 1991, Vol. 19, No. 7 1513-1520
© 1991
MOLECULAR BIOLOGY |
Identification and characterization of novel human endogenous retroviral sequences prefentially expressed in undifferentiated embryonal carcinoma cells
Department of Genetics, General and Molecular Biology, University of Naples Via Mezzocannone 8, 80124 Naples 1International Institute of Genetics and Biophysics, CNR Naples 2Institute of Medical Clinics I, University of Perugia 06100 Perugia, Italy
* To whom correspondence should be addressed
Received December 31, 1990. Revised March 13, 1991. Accepted March 13, 1991.
A novel endogenous retroviral sequence (ERV-9) has been isolated from a human embryonal carcinoma cDNA library by hybridization to a probe containing a recently described human repetitive element. DNA sequence analysis of the 4kb cDNA insert (pHE.1) revealed the presence of ORFs potentially coding for putative retrovirus-related gag, pol and env proteins. Northern blot and RNase protection experiments showed that RNA homologous to the pHE.1 insert is detected only in embryonal carcinoma cells as a 8 kb mRNA, and its expression is negatively regulated during retinoic acid induced differentiation of the human teratocarcinoma cell line NT2/D1. Using a pol specific probe we have isolated a genomic locus containing the ERV-9 sequences. Characterization by restriction enzyme analysis and DNA sequencing allowed us to define LTR-like sequences, that are composed by a complex array of subrepetitive elements. In addition we show that ERV-9 LTR sequences are capable to drive expression of linked CAT gene in a cell specific manner as LTR promoter activity has been detected only in NT2/D1 cells.
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