Guan Deyu, Altan-Bonnet Nihal, Parrott Andrew M, Arrigo Cindy J, Li Quan, Khaleduzzaman Mohammed, Li Hong, Lee Chee-Gun, Pe'ery Tsafi, Mathews Michael B
Departments of Biochemistry and Molecular Biology, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark, New Jersey 07103-1709, USA.
Mol Cell Biol. 2008 Jul;28(14):4629-41. doi: 10.1128/MCB.00120-08. Epub 2008 May 5.
Nuclear factor 90 (NF90) and its C-terminally extended isoform, NF110, have been isolated as DNA- and RNA-binding proteins together with the less-studied protein NF45. These complexes have been implicated in gene regulation, but little is known about their cellular roles and whether they are redundant or functionally distinct. We show that heterodimeric core complexes, NF90-NF45 and NF110-NF45, exist within larger complexes that are more labile and contain multiple NF90/110 isoforms and additional proteins. Depletion of the NF45 subunit by RNA interference is accompanied by a dramatic decrease in the levels of NF90 and NF110. Reciprocally, depletion of NF90 but not of NF110 greatly reduces the level of NF45. Coregulation of NF90 and NF45 is a posttranscriptional phenomenon, resulting from protein destabilization in the absence of partners. Depletion of NF90-NF45 complexes retards cell growth by inhibition of DNA synthesis. Giant multinucleated cells containing nuclei attached by constrictions accumulate when either NF45 or NF90, but not NF110, is depleted. This study identified NF45 as an unstable regulatory subunit of NF90-NF45 complexes and uncovered their critical role in normal cell division. Furthermore, the study revealed that NF90 is functionally distinct from NF110 and is more important for cell growth.
核因子90(NF90)及其C端延伸异构体NF110已作为DNA和RNA结合蛋白与研究较少的蛋白NF45一起被分离出来。这些复合物与基因调控有关,但对它们在细胞中的作用以及它们是冗余的还是功能不同的了解甚少。我们发现异二聚体核心复合物NF90-NF45和NF110-NF45存在于更大、更不稳定的复合物中,这些复合物包含多种NF90/110异构体和其他蛋白质。通过RNA干扰使NF45亚基缺失会伴随着NF90和NF110水平的显著下降。相反,NF90的缺失而非NF110的缺失会极大地降低NF45的水平。NF90和NF45的共同调节是一种转录后现象,是由于在没有伴侣的情况下蛋白质不稳定所致。NF90-NF45复合物的缺失通过抑制DNA合成来延缓细胞生长。当NF45或NF90(而非NF110)缺失时,会积累含有通过缢缩相连细胞核的巨大多核细胞。这项研究确定NF45是NF90-NF45复合物的不稳定调节亚基,并揭示了它们在正常细胞分裂中的关键作用。此外,该研究还表明NF90在功能上与NF110不同,并且对细胞生长更为重要。