Iouk T L, Aitchison J D, Maguire S, Wozniak R W
Department of Cell Biology, University of Alberta, Edmonton, Alberta, Canada T6G 2H7.
Mol Cell Biol. 2001 Feb;21(4):1260-71. doi: 10.1128/MCB.21.4.1260-1271.2001.
Ribosome biogenesis is regulated by environmental cues that coordinately modulate the synthesis of ribosomal components and their assembly into functional subunits. We have identified an essential yeast WD-repeat-containing protein, termed Rrb1p, that has a role in both the assembly of the 60S ribosomal subunits and the transcriptional regulation of ribosomal protein (RP) genes. Rrb1p is located in the nucleus and is concentrated in the nucleolus. Its presence is required to maintain normal cellular levels of 60S subunits, 80S ribosomes, and polyribosomes. The function of Rrb1p in ribosome biogenesis appears to be linked to its association with the ribosomal protein rpL3. Immunoprecipitation of Rrb1p from nuclear extracts revealed that it physically interacts with rpL3. Moreover, the overproduction of Rrb1p led to increases in cellular levels of free rpL3 that accumulated in the nucleus together with Rrb1p. The concentration of these proteins within the nucleus was dependent on ongoing protein translation. We also showed that overexpression of RRB1 led to an increase in the expression of RPL3 while all other examined RP genes were unaffected. In contrast, depletion of RRB1 caused an increase in the expression of all RP genes examined except RPL3. These results suggest that Rrb1p regulates RPL3 expression and uncouples it from the coordinated expression of other RP genes.
核糖体生物合成受环境信号调控,这些信号可协调调节核糖体组分的合成及其组装成功能亚基的过程。我们鉴定出一种必需的含WD重复序列的酵母蛋白,称为Rrb1p,它在60S核糖体亚基的组装和核糖体蛋白(RP)基因的转录调控中均发挥作用。Rrb1p位于细胞核中,且集中在核仁。其存在对于维持60S亚基、80S核糖体和多核糖体的正常细胞水平是必需的。Rrb1p在核糖体生物合成中的功能似乎与其与核糖体蛋白rpL3的关联有关。从核提取物中免疫沉淀Rrb1p表明它与rpL3存在物理相互作用。此外,Rrb1p的过量表达导致细胞核中与Rrb1p一起积累的游离rpL3细胞水平增加。这些蛋白质在细胞核内的浓度取决于正在进行的蛋白质翻译。我们还表明,RRB1的过表达导致RPL3的表达增加,而所有其他检测的RP基因均未受影响。相反,RRB1的缺失导致除RPL3外所有检测的RP基因的表达增加。这些结果表明,Rrb1p调节RPL3的表达,并使其与其他RP基因的协调表达解偶联。