Abstract
A chicken genomic library was screened with the human cDNA encoding the non-histone chromosomal protein HMG-17 and a 4565-base pair fragment containing the entire gene encoding this protein was isolated and characterized. Sequence analysis of the fragment revealed that from the start to end of transcription, the HMG-17 gene is 3293 base pairs long and is comprised of 6 exons ranging in size from 30 to 890 base pairs. Upstream of the putative cap site are both a CAAT box and a TATA box as well as several Sp1 binding sites. The gene has an extremely high content of G and C residues (75%) in a 1150-base pair fragment starting 500 base pairs from the putative cap site. This region satisfies the definition of an HpaII tiny fragment island. Southern analysis indicated that there is a single copy of this gene in chickens, whereas Northern analysis revealed that a single transcript is being synthesized from this gene. A comparison of the chicken and human cDNA and protein sequences and subsequent calculation of the evolutionary rates indicated that HMG-17 is a slowly evolving gene. The present article, which is the first study on the isolation and characterization of a complete gene coding for a high mobility group non-histone protein, indicates that the gene has features characteristic of housekeeping genes.
Highlights
A chickengenomiclibrary was screened with the exit of the DNA wrapped around the histone octamer, human cDNA encoding the non-histone chromosomal between the DNA and the histones [12]
Screening of the Chicken Genomic Library with Human HMG-17 putative regulatory role of these proteins was obtained from experiments which showed that microinjection of antibodies to HMG-17 into somaticcells inhibited transcription ( 5 ) and from reconstitution experiments which indicated that these proteins bind preferentially to salt-strippneudcleosomes containing transcribible sequences [3, 6]
We have reported that the nucleotide distribution in the human HMG-17 cDNAis somewhat unusual in that thoepen
Summary
Vol 263, No 8,Issue of March 15, pp. 3917-3923,1988 Printed in U.S.A. From the Laboratory of Molecular Carcinogenesis,National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892. Screening of the Chicken Genomic Library with Human HMG-17 putative regulatory role of these proteins was obtained from experiments which showed that microinjection of antibodies to HMG-17 into somaticcells inhibited transcription ( 5 ) and from reconstitution experiments which indicated that these proteins bind preferentially to salt-strippneudcleosomes containing transcribible sequences [3, 6]. The start of transcription is a purine and and 0.7 kb, the orderof which were determined by numerous in the HMG-17gene, initiation of transcription is at restriction digestions andSouthernanalyses (Fig. 1).Only the Aresidue a t position 869. Because the primer and subsequent identificationof the known HMG-17 peptide extension analysis revealed a single band which corresponds sequence [28], and by comparisontothecDNAsequence to the initiationof transcription at theA residue a t position isolated froma chicken expression librar(y29). 1500 I460 clcct#gc#g cgclgcctcc aaggggcrc-1 cctaagcggg agggggrcgg cccrttccgc CgggECtCgC gCtgCccccc tgccccgccg gcccgcgttg gggttg:agag agtccccccc
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