Mietz J A, Fewell J W, Kuff E L
Laboratory of Biochemistry, National Cancer Institute, Bethesda, Maryland 20892.
Mol Cell Biol. 1992 Jan;12(1):220-8. doi: 10.1128/mcb.12.1.220-228.1992.
Intracisternal A-particle (IAP) proviral elements are abundant and widely dispersed in the mouse genome. IAP-related transcripts have been detected in normal mouse tissues where expression is under genetic control. In this study, we sought to determine whether IAP expression in BALB/c thymus and lipopolysaccharide-stimulated B cells was due to selective or indiscriminate activation of IAP elements. cDNA libraries were prepared from each source. A total of 86 IAP cDNA clones were isolated from both libraries, and 37 of these were sequenced over a common 0.7- to 1.0-kb region of the IAP genome that included the 3' long terminal repeat (LTR). Three highly related families of elements were found to be expressed in the two cell types examined. All of the related elements had a distinctive U3 regulatory region. Thirteen individual IAP proviral elements were distinguished on the basis of sequence differences within the R region of the LTR. Hybridization of genomic DNA with element-specific oligonucleotide probes confirmed the presence of a restricted number of proviral copies in the lymphocyte-specific family of elements. Most of these copies were found to be methylated in the lymphocyte DNA, but at least seven were hypomethylated in their 5' LTRs. This study shows that activation of IAP elements in normal normal mouse lymphocytes is highly selective. Activation is probably a function of both sequence specificity and methylation status of the proviral LTR.
脑池内A颗粒(IAP)前病毒元件在小鼠基因组中含量丰富且广泛分布。在受基因控制表达的正常小鼠组织中已检测到IAP相关转录本。在本研究中,我们试图确定BALB/c胸腺和脂多糖刺激的B细胞中IAP的表达是由于IAP元件的选择性激活还是非特异性激活。从每个来源制备cDNA文库。从两个文库中总共分离出86个IAP cDNA克隆,其中37个在IAP基因组一个常见的0.7至1.0 kb区域进行测序,该区域包括3'长末端重复序列(LTR)。在检测的两种细胞类型中发现有三个高度相关的元件家族表达。所有相关元件都有一个独特的U3调控区。根据LTR的R区域内的序列差异区分出13个单独的IAP前病毒元件。用元件特异性寡核苷酸探针与基因组DNA杂交证实,淋巴细胞特异性元件家族中存在数量有限的前病毒拷贝。这些拷贝中的大多数在淋巴细胞DNA中被甲基化,但至少有七个在其5' LTR中是低甲基化的。这项研究表明,正常小鼠淋巴细胞中IAP元件的激活具有高度选择性。激活可能是前病毒LTR的序列特异性和甲基化状态共同作用的结果。