Iwata Y, Shinomura T, Kurita K, Zako M, Kimata K
Institute for Molecular Science of Medicine, Aichi Medical University, Nagakute, Aichi 480-11, Japan.
Biochem J. 1998 May 1;331 ( Pt 3)(Pt 3):959-64. doi: 10.1042/bj3310959.
PG-Lb was originally characterized as a small chondroitin/dermatan sulphate proteoglycan expressed preferentially in the zones of flattened chondrocytes in developing chick limb cartilage. The occurrence of this proteoglycan in mammalian cartilage has been shown by the isolation of a cDNA clone from mouse cartilage cDNA library [Kurita, Shinomura,Ujita, Zako, Kida, Iwata and Kimata (1996) Biochem. J. 318, 909-914]. To understand the regulation mechanisms for such a unique expression, we have investigated a genomic DNA structure of the PG-Lb gene. The gene is composed of seven exons and six introns spanning more than 50 kb. The leucine-rich repeats are encoded from exon V to exon VII. The transcription initiation site has been determined by rapid amplification of the cDNA ends ('5'-RACE'). The possible TATA box was detected about 90 bp upstream of the adenosine residue that was numbered as position +1. Further analyses of 1.5 kb of the 5' flanking region and 2.2 kb of the first intron have revealed several potential binding motifs for transcription factors such as Sox 5 and 9. The presence of those sequences in the PG-Lb gene was discussed in relation to the unique expression of this proteoglycan. The chromosomal localization of the murine PG-Lb gene was determined to be on the mouse chromosome 10 by the fluorescence-in-situ-hybridization ('FISH') method.
PG-Lb最初被鉴定为一种小的硫酸软骨素/硫酸皮肤素蛋白聚糖,在发育中的鸡肢体软骨中扁平软骨细胞区域优先表达。从小鼠软骨cDNA文库中分离出一个cDNA克隆,证明了这种蛋白聚糖在哺乳动物软骨中的存在[栗田、筱村、宇田、迫子、木田、岩田和木又田(1996年)《生物化学杂志》318卷,909 - 914页]。为了了解这种独特表达的调控机制,我们研究了PG-Lb基因的基因组DNA结构。该基因由七个外显子和六个内含子组成,跨度超过50kb。富含亮氨酸的重复序列由外显子V到外显子VII编码。转录起始位点已通过cDNA末端快速扩增(“5'-RACE”)确定。在编号为+1的腺苷残基上游约90bp处检测到可能的TATA框。对1.5kb的5'侧翼区域和2.2kb的第一个内含子的进一步分析揭示了几个转录因子如Sox 5和9的潜在结合基序。结合这种蛋白聚糖的独特表达讨论了PG-Lb基因中这些序列的存在。通过荧光原位杂交(“FISH”)方法确定小鼠PG-Lb基因的染色体定位在小鼠10号染色体上。