Further characterization of the murine collagenase (type IVB) gene promoter and analysis of mRNA expression in murine tissues.
The collagenase B type IV (Col4B) gene is highly expressed in the osteoclast, the primary bone-resorbing cell. However, factors that regulate expression of the Col4B gene are not well characterized. A murine P1 genomic clone containing a 94 kb sequence insert which contains the Col4B gene was isolated. A 4 kb EcoR1 DNA fragment containing the 5' flanking sequence of the gene was further subcloned and restriction mapped. Putative transcription factors such as SRY, Lyf-1, and GATA1 and 2, binding motifs were identified by sequence analysis in this promoter region. Enhancer and suppressor regions were mapped by transient expression of Col4B gene promoter deletion mutant-luciferase reporter gene constructs in HepG2 cells. Col4B mRNA expression in different murine tissues was analyzed by reverse transcription-polymerase chain reaction and demonstrated high levels of expression in bone, clavaria, spleen and thymus. This promoter provides a valuable tool for targeting gene expression to the osteoclast.
Roach
J
J
Department of Medicine and Hematology, University of Texas Health Science Center, San Antonio 78284-7880, USA.
Choi
S J
SJ
Schaub
R L
RL
Leach
R J
RJ
Roodman
G D
GD
Reddy
S V
SV
D000818:Animals; D001483:Base Sequence; D017364:Collagenases / Q000096:biosynthesis* / Q000235:genetics*; D017930:Genes, Reporter; D008156:Luciferases / Q000096:biosynthesis; D051379:Mice / Q000235:genetics*; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010010:Osteoclasts / Q000201:enzymology; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000096:biosynthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D017384:Sequence Deletion; D014158:Transcription, Genetic
D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D008156:Luciferases; D017364:Collagenases