Cohen Merav, Santarella Rachel, Wiesel Naama, Mattaj Iain, Gruenbaum Yosef
Division of Cell Biology, MRC-Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
Methods Cell Biol. 2008;88:411-29. doi: 10.1016/S0091-679X(08)00421-4.
The nuclear lamina is found between the inner nuclear membrane and the peripheral chromatin. Lamins are the main components of the nuclear lamina, where they form protein complexes with integral proteins of the inner nuclear membrane, transcriptional regulators, histones and chromatin modifiers. Lamins are required for mechanical stability, chromatin organization, Pol II transcription, DNA replication, nuclear assembly, and nuclear positioning. Mutations in human lamins cause at least 13 distinct human diseases, collectively termed laminopathies, affecting muscle, adipose, bone, nerve and skin cells, and range from muscular dystrophies to accelerated aging. Caenorhabditis elegans has unique advantages in studying lamins and nuclear lamina genes including low complexity of lamina genes and the unique ability of bacterially expressed C. elegans lamin protein to form stable 10 nm fibers. In addition, transgenic techniques, simple application of RNA interference, sophisticated genetic analyses, and the production of a large collection of mutant lines, all make C. elegans especially attractive for studying the functions of its nuclear lamina genes. In this chapter we will include a short review of our current knowledge of nuclear lamina in C. elegans and will describe electron microscopy techniques used for their analyses.
核纤层位于内核膜和外周染色质之间。核纤层蛋白是核纤层的主要成分,它们在内核膜的整合蛋白、转录调节因子、组蛋白和染色质修饰因子之间形成蛋白质复合物。核纤层蛋白对于机械稳定性、染色质组织、RNA聚合酶II转录、DNA复制、核组装和核定位是必需的。人类核纤层蛋白的突变会导致至少13种不同的人类疾病,统称为核纤层病,影响肌肉、脂肪、骨骼、神经和皮肤细胞,范围从肌肉萎缩症到加速衰老。秀丽隐杆线虫在研究核纤层蛋白和核纤层基因方面具有独特优势,包括核纤层基因的低复杂性以及细菌表达的秀丽隐杆线虫核纤层蛋白形成稳定的10纳米纤维的独特能力。此外,转基因技术、RNA干扰的简单应用、复杂的遗传分析以及大量突变株系的产生,都使得秀丽隐杆线虫在研究其核纤层基因功能方面特别有吸引力。在本章中,我们将简要回顾目前我们对秀丽隐杆线虫核纤层的了解,并描述用于其分析的电子显微镜技术。