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J. Biol. Chem., Vol. 265, Issue 25, 14922-14931, 09, 1990
M Srivastava, OW McBride, PJ Fleming, HB Pollard and AL Burns
Nucleolin, a eukaryotic nucleolar phosphoprotein, is involved in the
synthesis and maturation of ribosomes. To characterize the genomic
organization and regulatory sequences of this gene, two overlapping lambda
clones containing the human nucleolin gene plus flanking regions were
isolated from a genomic library using human nucleolin cDNA. Southern blots
of genomic DNA from human, several mammals, chicken, and yeast revealed
that the nucleolin gene is well conserved across these species. The gene
consists of 14 exons with 13 intervening sequences and spans approximately
11 kilobases of DNA. Analysis of the splice junctions indicated that the
amino-terminal domain and the four RNA binding domains plus the nuclear
localization signal are split into adjacent exons. Sequences from the
5'-flanking and the first intron contain a high content of GC residues
which is consistent with nucleolin being a "housekeeping" gene. Promoter
elements include an atypical TATA box (GTTA), one CCAAT box much further
from the initiation site, three reverse compliments of CCAAT (ATTGG), and
two pyrimidine-rich nucleotide stretches. In addition, this region and the
first intron contain numerous potential Sp1, GCF, CRE-fos, GCN, AP-1, AP-2,
UCE, and sequences similar to the glucocorticoid receptor binding site. The
transcription start site was determined by primer extension and S1 nuclease
mapping of RNA from human liver. One Kpn and three Alu repeats were found
within two of the middle introns. The 3'- untranslated portion of the gene
contains five homology blocks in a 100- base pair region that are highly
conserved among human, mouse, and hamster genomes. Finally, we have
determined that the human nucleolin gene is located on chromosome 2q12-qter
and is present at one copy per haploid genome. A restriction fragment
length polymorphism with EcoRI has been detected in the gene.
Genomic organization and chromosomal localization of the human nucleolin gene
Cardiorenal Division, Food and Drug Administration, Bethesda, Maryland.
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