A novel karyopherin-β homolog is developmentally and hormonally regulated in fetal lung
A novel karyopherin-β homolog is developmentally and hormonally regulated in fetal lung
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DOI：
10.1165/ajrcmb.22.4.3929
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发表时间：
2000-04-01
影响因子：
6.4
通讯作者：
Kaplan, F
中科院分区：
文献类型：
--
作者：
Zhang, CD;Sweezey, NB;Kaplan, F
为了研究肺器官发生的分子机制，我们采用代表性差异分析在胎鼠肺发育模型中寻找糖皮质激素诱导基因。从胎儿和成年大鼠肺中制备的信使RNA用于制备“代表性扩增子”，利用成人肺互补DNA （cDNA）扩增子作为“驱动器”，在连续的减法杂交/扩增中分离出目标胎儿肺特异性cDNA，以一个保守且与8个人类/啮齿类动物表达的序列标签具有近乎完美同源性的单克隆为模板，进行cDNA末端的5‘和3’快速扩增和SPICE （system for polymerase chain reaction amplification of cDNA ends）反应，获得3.6 kb cDNA, LGL2 (Genbank，AF 110195)编码963个氨基酸的推导多肽（lgl2）。Northern分析证实LGL2在胎儿肺中存在差异表达（最大表达期为假腺期，妊娠14 ~ 16天），由糖皮质激素诱导，上皮相对于间质富集。在妊娠第16周的人胎儿肺以及人和大鼠胎儿脑、心、肠和肾中也检测到LGL2。我们将LGL2定位到染色体1p33-34.2。与基因组数据库的序列比较，lgl2是核输入蛋白核丝蛋白- β家族的成员，与转运蛋白sr的同源性最大。lgl2在假腺发育阶段的最大表达与胎儿肺器官发生中调节重要信号转导通路的关键转录因子的最大表达相协调。我们提出lgl2在胎儿肺发育过程中调节信号转导的转录因子的核输入中的作用。
To investigate molecular mechanisms of lung organogenesis, we used representational difference analysis to search for glucocorticoid-inducible genes in developing lung in a fetal rat model. Messenger RNA prepared from fetal and adult rat lung was used to prepare "representative amplicons," Adult-lung complementary DNA (cDNA) amplicons were used as "driver" in successive rounds of subtractive hybridization/amplification to isolate target fetal lung-specific cDNAs, A single clone, which was conserved and had near-perfect homology to eight human/rodent expressed sequence tags, was used as template for 5' and 3' rapid amplification of cDNA ends and SPICE (system for polymerase chain reaction amplification of cDNA ends) reactions to obtain the 3.6-kb cDNA, LGL2 (Genbank, AF 110195) encoding a deduced polypeptide (lgl2) of 963 amino acids. Northern analysis confirmed that LGL2 is differentially expressed in fetal lung (maximal during the pseudoglandular stage, gestational Days 14 to 16), induced by glucocorticoid, and enriched in epithelium relative to the mesenchyme. LGL2 was also detected in human fetal lung at gestational Week 16 as well as in human and rat fetal brain, heart, intestine, and kidney. We mapped LGL2 to chromosome 1p33-34.2. Comparison with sequences in the genome database identified lgl2 as a member of the karyopherin-beta family of nuclear import proteins, with greatest homology to transportin SR. Maximal expression of LGL2 in the pseudoglandular stage of development is coordinate with that of key transcription factors that regulate prominent signal transduction pathways in fetal lung organogenesis, We propose a role for lgl2 in nuclear import of transcription factors that regulate signal transduction during fetal lung development.