Blewett Nathan H, Iben James R, Gaidamakov Sergei, Maraia Richard J
Intramural Research Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Rockville, Maryland, USA.
Commissioned Corps, U.S. Public Health Service, Rockville, Maryland, USA
Mol Cell Biol. 2017 May 2;37(10). doi: 10.1128/MCB.00588-16. Print 2017 May 15.
Human La antigen (Sjögren's syndrome antigen B [SSB]) is an abundant multifunctional RNA-binding protein. In the nucleoplasm, La binds to and protects from 3' exonucleases, the ends of precursor tRNAs, and other transcripts synthesized by RNA polymerase III and facilitates their maturation, while a nucleolar isoform has been implicated in rRNA biogenesis by multiple independent lines of evidence. We showed previously that conditional La knockout (La cKO) from mouse cortex neurons results in defective tRNA processing, although the pathway(s) involved in neuronal loss thereafter was unknown. Here, we demonstrate that La is stably associated with a spliced pre-tRNA intermediate. Microscopic evidence of aberrant nuclear accumulation of 5.8S rRNA in La cKO is supported by a 10-fold increase in a pre-5.8S rRNA intermediate. To identify pathways involved in subsequent neurodegeneration and loss of brain mass in the cKO cortex, we employed mRNA sequencing (mRNA-Seq), immunohistochemistry, and other approaches. This revealed robust enrichment of immune and astrocyte reactivity in La cKO cortex. Immunohistochemistry, including temporal analyses, demonstrated neurodegeneration, followed by astrocyte invasion associated with immune response and decreasing cKO cortex size over time. Thus, deletion of La from postmitotic neurons results in defective pre-tRNA and pre-rRNA processing and progressive neurodegeneration with loss of cortical brain mass.
人La抗原(干燥综合征抗原B [SSB])是一种丰富的多功能RNA结合蛋白。在核质中,La与前体tRNA的末端以及RNA聚合酶III合成的其他转录本结合,并保护它们免受3'外切核酸酶的降解,促进其成熟,而一种核仁异构体已被多条独立证据证明与rRNA生物合成有关。我们之前表明,从小鼠皮质神经元中条件性敲除La(La cKO)会导致tRNA加工缺陷,尽管此后神经元丢失所涉及的途径尚不清楚。在这里,我们证明La与剪接的前体tRNA中间体稳定相关。在La cKO中5.8S rRNA异常核积累的显微镜证据得到了前体5.8S rRNA中间体增加10倍的支持。为了确定cKO皮质中随后神经退行性变和脑质量丧失所涉及的途径,我们采用了mRNA测序(mRNA-Seq)、免疫组织化学和其他方法。这揭示了La cKO皮质中免疫和星形胶质细胞反应性的强烈富集。包括时间分析在内的免疫组织化学显示了神经退行性变,随后是与免疫反应相关的星形胶质细胞侵入以及随着时间推移cKO皮质大小的减小。因此,有丝分裂后神经元中La的缺失导致前体tRNA和前体rRNA加工缺陷以及进行性神经退行性变和皮质脑质量丧失。