PMID- 10406845
OWN - NLM
STAT- MEDLINE
DCOM- 19991014
LR  - 20190513
IS  - 0959-6658 (Print)
IS  - 0959-6658 (Linking)
VI  - 9
IP  - 8
DP  - 1999 Aug
TI  - Genomic organization and promoter activity of glucosidase I gene.
PG  - 797-806
AB  - Glucosidase I initiates the processing of asparagine (N-) linked glycoproteins by
      removing the distal alpha1,2-linked glucosyl residue of the tetradecasaccharide
      Glc(3)Man(9)GlcNAc(2). The gene encoding this enzyme was isolated and its
      structural organization and promoter activity determined. The major transcript
      for glucosidase I on northern blot appeared to be 3.1 kb; Southern blotting and
      DNA sequencing indicated the size of the gene to be 6.8 kb, comprising four exons
      separated by three introns. The first exon encodes the cytoplasmic tail and
      transmembrane domain; the fourth encodes the putative catalytic domain of the
      enzyme. Exon-intron junctions are flanked by consensus splice donor and acceptor 
      sequences. Transcription initiation sites were mapped by primer extension,
      ribonuclease protection assay and RT-PCR analysis. Primer extension results
      showed multiple initiation sites at -150, -156, and -272 bp relative to the
      translation initiation codon ATG. Sequence analysis of 5' flanking region showed 
      no canonical TATA box, a high GC content, Sp1 and ETF binding sites (typical of a
      housekeeping gene promoter). Also noteworthy, the promoter region contains
      several generic STAT factor binding sites, one nearly perfect, and two half GR
      binding elements. Other cis- acting elements recognized by transcription factors 
      such as AP-2, NF-kappaB, estrogen receptor, and progesterone receptor (PR) were
      also present in the putative promoter region. To determine the promoter activity,
      a construct encompassing the region between -2114 to -5 bp of the putative
      promoter was ligated to the chloramphenicol acetyltransferase (CAT) reporter
      plasmid and transiently transfected into COS 7 cells. CAT assay results clearly
      show transcriptional activity of the promoter.
FAU - Khan, F A
AU  - Khan FA
AD  - Department of Animal and Avian Sciences, University of Maryland, College Park, MD
      20742-2311, USA.
FAU - Varma, G M
AU  - Varma GM
FAU - Vijay, I K
AU  - Vijay IK
LA  - eng
GR  - DK 19682/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Glycobiology
JT  - Glycobiology
JID - 9104124
RN  - 0 (Polysaccharides)
RN  - 0 (Recombinant Fusion Proteins)
RN  - EC 2.3.1.28 (Chloramphenicol O-Acetyltransferase)
RN  - EC 3.2.1.- (glucosidase I)
RN  - EC 3.2.1.20 (alpha-Glucosidases)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - COS Cells
MH  - Carbohydrate Sequence
MH  - Chloramphenicol O-Acetyltransferase/genetics
MH  - Exons
MH  - Female
MH  - Genes, Reporter
MH  - Genomic Library
MH  - Humans
MH  - Introns
MH  - Mice
MH  - Molecular Sequence Data
MH  - Organ Specificity
MH  - Polysaccharides/chemistry/metabolism
MH  - *Promoter Regions, Genetic
MH  - Recombinant Fusion Proteins/biosynthesis
MH  - Substrate Specificity
MH  - *Transcription, Genetic
MH  - Transfection
MH  - alpha-Glucosidases/*genetics/metabolism
EDAT- 1999/07/16 00:00
MHDA- 1999/07/16 00:01
CRDT- 1999/07/16 00:00
PHST- 1999/07/16 00:00 [pubmed]
PHST- 1999/07/16 00:01 [medline]
PHST- 1999/07/16 00:00 [entrez]
AID - cwc074 [pii]
AID - 10.1093/glycob/9.8.797 [doi]
PST - ppublish
SO  - Glycobiology. 1999 Aug;9(8):797-806. doi: 10.1093/glycob/9.8.797.