The human gC1qR/p32 gene, C1qBP. Genomic organization and promoter analysis.
gC1qR is an ubiquitously expressed cell protein that interacts with the globular heads of C1q (gC1q) and many other ligands. In this study, the 7.8-kilobase pair (kb) human gC1qR/p32 (C1qBP) gene was cloned and found to consist of 6 exons and 5 introns. Analysis of a 1.3-kb DNA fragment at the 5'-flanking region of this gene revealed the presence of multiple TATA, CCAAT, and Sp1 binding sites. Luciferase reporter assays performed in different human cell lines demonstrated that the reporter gene was ubiquitously driven by this 1.3-kb fragment. Subsequent 5' and 3' deletion of this fragment confined promoter elements to within 400 base pairs (bp) upstream of the translational start site. Because the removal of the 8-bp consensus TATATATA at -399 to -406 and CCAAT at -410 to -414 did not significantly affect the transcription efficiency of the promoter, GC-rich sequences between this TATA box and the translation start site may be very important for the promoter activity of the C1qBP gene. One of seven GC-rich sequences in this region binds specifically to PANC-1 nuclear extracts, and the transcription factor Sp1 was shown to bind to this GC-rich sequence by the supershift assay. Primer extension analysis mapped three major transcription start regions. The farthest transcription start site is 49 bp upstream of the ATG translation initiation codon and is in close proximity of the specific SP1 binding site.
Department of Zoology, University of Hong Kong, Pokfulam, Hong Kong, China.
A J
AJ
Tye
B
B
Ghebrehiwet
N
N
Guo
K N
KN
Sastry
B K
BK
Chow
E I
EI
Peerschke
B L
BL
Lim
D020413:3' Untranslated Regions / Q000235:genetics; D020121:5' Untranslated Regions / Q000235:genetics; D001483:Base Sequence; D002352:Carrier Proteins; D002460:Cell Line; D015922:Complement C1q / Q000378:metabolism; D016384:Consensus Sequence; D005091:Exons; D015698:Genomic Library; D006801:Humans; D018960:Hyaluronan Receptors; D007438:Introns; D008562:Membrane Glycoproteins; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D011951:Receptors, Complement / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D015183:Restriction Mapping; D017384:Sequence Deletion; D016385:TATA Box; D014162:Transfection
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; C087299:C1QBP protein, human; D002352:Carrier Proteins; D018960:Hyaluronan Receptors; D008562:Membrane Glycoproteins; D024101:Mitochondrial Proteins; D011951:Receptors, Complement; D011994:Recombinant Proteins; C032402:complement 1q receptor; D015922:Complement C1q