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. 2009 Jun 18;583(12):2049-53.
doi: 10.1016/j.febslet.2009.05.023. Epub 2009 May 18.

Posttranscriptional down-regulation of small ribosomal subunit proteins correlates with reduction of 18S rRNA in RPS19 deficiency

Jitendra Badhai  1 , Anne-Sophie Fröjmark, Hamid Reza Razzaghian, Edward Davey, Jens Schuster, Niklas Dahl
Affiliations

Posttranscriptional down-regulation of small ribosomal subunit proteins correlates with reduction of 18S rRNA in RPS19 deficiency

Jitendra Badhai et al. FEBS Lett. .

Abstract

Ribosomal protein S19 (RPS19) is mutated in patients with Diamond-Blackfan anemia (DBA). We hypothesized that decreased levels of RPS19 lead to a coordinated down-regulation of other ribosomal (r-)proteins at the subunit level. We show that small interfering RNA (siRNA) knock-down of RPS19 results in a relative decrease of small subunit (SSU) r-proteins (S20, S21 and S24) when compared to large subunit (LSU) r-proteins (L3, L9, L30 and L38) VSports手机版. This correlates with a relative decrease in 18S rRNA with respect to 28S rRNA. The r-protein mRNA levels remain relatively unchanged indicating a post transcriptional regulation of r-proteins at the level of subunit formation. .

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Figures

Figure 1
Figure 1
Levels of r-proteins after siRNA induced knock-down of RPS19 in TF-1-B cells. Western blot analysis of induced (dox+) and non-induced (dox-) TF-1-B cells, respectively, using the primary antibodies targeting r-proteins, β-actin and fibrillarin. Doxycycline induction results in a reduction of S19 r-protein levels to 30% of levels in non-induced cells (S; small subunit protein: L; large subunit protein. Pictures of the complete Western blots are presented in Supplementary Figure 1).
Figure 2
Figure 2
Levels of mRNA and protein corresponding to seven r-proteins and fibrillarin (Fib) after siRNA mediated knock-down of RPS19 (7 days of induction). Three SSU r-proteins (S) and four LSU proteins (L) are analyzed. (A) Amounts of mRNA levels in doxycycline induced TF-1-B cells related to the amounts in non-induced cells (set to 1.0). A significant down regulation was observed for RPL3 mRNA only (* denotes p<0.05; three independent experiments). Quantification was performed by qRT/PCR (primer sequences are available upon request) and normalized to β-actin. (B) Amounts of r-protein in doxycycline induced TF-1-B cells related to the amounts in untreated cells (set to 1.0) as determined by western blot analysis using specific antibodies. A significant reduction was observed for the three SSU r-proteins (* denotes p<0.05; three independent experiments) but not for any of the LSU r-proteins using two tailed student’s t-test. (C) Ratio of pooled SSU r-protein levels to pooled LSU r-protein levels in non-induced TF-1-B cells (set to 1,0) and induced TF-1-B cells, respectively (* denotes p<0.05; two tailed student’s t-test). The levels of individual r-proteins are normalized to β-actin and averaged before pooling.
Figure 3
Figure 3
Amounts of 18S rRNA to S28 rRNA in TF-1-B cells after siRNA mediated knock-down of RPS19. Total RNA was isolated from doxycycline induced and uninduced TF-1-B cells after 2, 4 and 7 days, respectively. The amounts of rRNAs were quantified using the Agilent 2100 bioanalyser. (A) The 18S to 28S rRNA ratio in induced cells as compared to uninduced cells set to 1 (* denotes p<0.05; two tailed student’s t-test). (B) The mean values of the 18S to 28S ratio in uninduced (dox-) and induced (dox+) cells at day 2, 4 and 7 with standard deviations. The mean values at each day is based on three different experiments.
Figure 4
Figure 4
Ratio of pooled SSU r-protein levels to pooled LSU r-protein levels in lymphoblastoid cell lines (LCLs) from normal controls and RPS19 deficient DBA patients. The ratio in LCLs from healthy controls (left) is significantly higher than that in DBA patients with RPS19 mutations (right; * denote p<0.05). The levels of individual r-proteins are normalized to β-actin and averaged before pooling. The ratio in controls is set to 1.0.
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References

    1. Steitz TA, Moore PB. RNA, the first macromolecular catalyst: the ribosome is a ribozyme. Trends Biochem Sci. 2003;28:411–418. - PubMed
    1. Ellis SR, Lipton JM. Diamond Blackfan anemia: a disorder of red blood cell development. Curr Top Dev Biol. 2008;82:217–241. - PubMed
    1. Draptchinskaia N. The gene encoding ribosomal protein S19 is mutated in Diamond-Blackfan anaemia. Nat Genet. 1999;21:169–175. - PubMed
    1. Gazda HT. Ribosomal protein S24 gene is mutated in Diamond-Blackfan anemia. Am J Hum Genet. 2006;79:1110–1118. - PMC (VSports在线直播) - PubMed
    1. Cmejla R, Cmejlova J, Handrkova H, Petrak J, Pospisilova D. Ribosomal protein S17 gene (RPS17) is mutated in Diamond-Blackfan anemia. Hum Mutat. 2007;28:1178–1182. - "V体育平台登录" PubMed

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