Identification of proteins crosslinked to RNA in 40S ribosomal subunits of Saccharomyces cerevisiae

Xiang, Rui; Lee, John C.

doi:10.1016/0300-9084(89)90024-2

Cited by 3 publications

(3 citation statements)

References 27 publications

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“…Indeed, we have identified RPS13 and RPS18 associated with a large U3snoRNP complex containing nucleolin-like protein (our unpublished data), which is involved in early processing of pre-rRNA in B. oleracea (Saez-Vasquez et al, 2004). Moreover, it has been also shown that these two RP proteins bind 18S rRNA and are 40S early ribosome assembly proteins in yeast (Xiang and Lee, 1989). Nevertheless, despite all these observations, it is likely that the phenotype observed in Atnuc-L1 can also be due to an effect on one or several of the multifunctional roles described for nucleolin in eukaryotes (Ginisty et al, 1999;Srivastava and Pollard, 1999).…”

Section: Discussionmentioning

confidence: 78%

Characterization of AtNUC-L1 Reveals a Central Role of Nucleolin in Nucleolus Organization and Silencing of AtNUC-L2 Gene in Arabidopsis

Pontvianne

Matı́a

Douet

et al. 2007

MBoC

125

207

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Nucleolin is one of the most abundant protein in the nucleolus and is a multifunctional protein involved in different steps of ribosome biogenesis. In contrast to animals and yeast, the genome of the model plant Arabidopsis thaliana encodes two nucleolin-like proteins, AtNUC-L1 and AtNUC-L2. However, only the AtNUC-L1 gene is ubiquitously expressed in normal growth conditions. Disruption of this AtNUC-L1 gene leads to severe plant growth and development defects. AtNUC-L1 is localized in the nucleolus, mainly in the dense fibrillar component. Absence of this protein in Atnuc-L1 plants induces nucleolar disorganization, nucleolus organizer region decondensation, and affects the accumulation levels of pre-rRNA precursors. Remarkably, in Atnuc-L1 plants the AtNUC-L2 gene is activated, suggesting that AtNUC-L2 might rescue, at least partially, the loss of AtNUC-L1. This work is the first description of a higher eukaryotic organism with a disrupted nucleolin-like gene and defines a new role for nucleolin in nucleolus structure and rDNA chromatin organization.

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Section: Discussionmentioning

confidence: 78%

Characterization of AtNUC-L1 Reveals a Central Role of Nucleolin in Nucleolus Organization and Silencing of AtNUC-L2 Gene in Arabidopsis

Pontvianne

Matı́a

Douet

et al. 2007

MBoC

125

207

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“…In the Atnuc-l1 mutant, pre-rRNA precursor accumulation is decreased, and primary cleavage and subsequent pre-rRNA processing are disrupted (Kojima et al, 2007;Petricka and Nelson, 2007;Pontvianne et al, 2007). In yeast, the RPL23a ortholog is required for efficient pre-rRNA processing (van Beekvelt et al, 2001), and both RPS13 and RPS18 are universally conserved, core structural components of the SSU that bind 18S rRNA and are associated with the prerRNA processing complex (Xiang and Lee, 1989;Yusupov et al, 2001;Brodersen et al, 2002;Pontvianne et al, 2007). The striking similarity of phenotype between rpl23aa, rps5b, rps13b, rps18a, rpl24b, and Atnuc-l1 mutants may thus represent the in vivo response to decreased production of processed rRNAs and the concomitant effects on ribosome biogenesis and protein production, with the observed severity gradient being a measure of the extent of the reduction in mature rRNAs.…”

Section: Discussionmentioning

confidence: 99%

Arabidopsis Ribosomal Proteins RPL23aA and RPL23aB Are Differentially Targeted to the Nucleolus and Are Disparately Required for Normal Development

2008

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Protein synthesis is catalyzed by the ribosome, a two-subunit enzyme comprised of four ribosomal RNAs and, in Arabidopsis (Arabidopsis thaliana), 81 ribosomal proteins (r-proteins). Plant r-protein genes exist as families of multiple expressed members, yet only one r-protein from each family is incorporated into any given ribosome, suggesting that many r-protein genes may be functionally redundant or development/tissue/stress specific. Here, we characterized the localization and gene-silencing phenotypes of a large subunit r-protein family, RPL23a, containing two expressed genes (RPL23aA and RPL23aB). Live cell imaging of RPL23aA and RPL23aB in tobacco with a C-terminal fluorescent-protein tag demonstrated that both isoforms accumulated in the nucleolus; however, only RPL23aA was targeted to the nucleolus with an N-terminal fluorescent protein tag, suggesting divergence in targeting efficiency of localization signals. Independent knockdowns of endogenous RPL23aA and RPL23aB transcript levels using RNA interference determined that an RPL23aB knockdown did not alter plant growth or development. Conversely, a knockdown of RPL23aA produced a pleiotropic phenotype characterized by growth retardation, irregular leaf and root morphology, abnormal phyllotaxy and vasculature, and loss of apical dominance. Comparison to other mutants suggests that the phenotype results from reduced ribosome biogenesis, and we postulate a link between biogenesis, microRNA-target degradation, and maintenance of auxin homeostasis. An additional RNA interference construct that coordinately silenced both RPL23aA and RPL23aB demonstrated that this family is essential for viability.

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“…Marczinovits and Molnar (1982) used UV cross-linking to study RNA-protein interactions in the 30S subunits of rat liver hnRNP. Xiang and Lee (1989) identified proteins cross-linked to RNA in the 40S ribosomal subunits of Saccharomyces cerevisiae. Leibold and Munro (1988) found a cytoplasmic protein bound to the 5′ UTR of ferritin mRNAs.…”

Section: Commentary Background Informationmentioning

confidence: 99%

Identification of RNA Binding Proteins by UV Cross‐Linking

Luo

Reed

2003

CP Molecular Biology

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One of the major focuses of modern biology is to understand the dynamics of RNA-protein interactions, including factors that interact with mRNA, rRNA, tRNA, snRNA, hnRNA, siRNA, and viral RNA. Identification the direct interactions between proteins and RNA has greatly advanced our knowledge about the function of RNA-protein machines. UV crosslinking is a straightforward and powerful tool in this endeavor.

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Identification of proteins crosslinked to RNA in 40S ribosomal subunits of Saccharomyces cerevisiae

Cited by 3 publications

References 27 publications

Characterization of AtNUC-L1 Reveals a Central Role of Nucleolin in Nucleolus Organization and Silencing of AtNUC-L2 Gene in Arabidopsis

Characterization of AtNUC-L1 Reveals a Central Role of Nucleolin in Nucleolus Organization and Silencing of AtNUC-L2 Gene in Arabidopsis

Arabidopsis Ribosomal Proteins RPL23aA and RPL23aB Are Differentially Targeted to the Nucleolus and Are Disparately Required for Normal Development

Identification of RNA Binding Proteins by UV Cross‐Linking

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