Pol5 is an essential ribosome biogenesis factor required for 60S ribosomal subunit maturation in Saccharomyces cerevisiae

Abstract

In Saccharomyces cerevisiae, more than 250 trans-acting factors are involved in the maturation of 40S and 60S ribosomal subunits. The expression of most of these factors is transcriptionally coregulated to ensure correct ribosome production under a wide variety of environmental and intracellular conditions. Here, we identified the essential nucleolar Pol5 protein as a novel trans-acting factor required for the synthesis of 60S ribosomal subunits. Pol5 weakly and/or transiently associates with early to medium pre-60S ribosomal particles. Depletion of and temperature-sensitive mutations in Pol5 result in a deficiency of 60S ribosomal subunits and accumulation of half-mer polysomes. Both processing of 27SB pre-rRNA to mature 25S rRNA and release of pre-60S ribosomal particles from the nucle(ol)us to the cytoplasm are impaired in the Pol5depleted strain. Moreover, we identified the genes encoding ribosomal proteins uL23 and eL27A as multicopy suppressors of the slow growth of a temperature-sensitive pol5 mutant. These results suggest that Pol5 could function in ensuring the correct folding of 25S rRNA domain III; thus, favoring the correct assembly of these two ribosomal proteins at their respective binding sites into medium pre-60S ribosomal particles. Pol5 is homologous to the human tumor suppressor Myb-binding protein 1A (MYBBP1A). However, expression of MYBBP1A failed to complement the lethal phenotype of a pol5 null mutant strain though interfered with 60S ribosomal subunit biogenesis.

TheMybfamily genes in the rice pathogenMagnaporthe oryzae: Finding and deleting more family members involved in pathogenicity

“…That indicates that the genes have diverse functions (Hogues et al ., 2008; Yu et al ., 2010; Rodríguez-Sánchez et al ., 2011; Mukherjee et al ., 2012; Pattabiraman and Gonda, 2013; Roy, 2016; X. Wang et al ., 2018; Ma and Constabel, 2019; Ramos-Sáenz et al ., 2019), and might not all be traditional transcription factors. The latter is often implied for plant Myb proteins (Wieser and Adams, 1995; Mateos et al ., 2006; Du et al ., 2009; Dubos et al ., 2010; Lin et al ., 2011, 2012; Prouse and Campbell, 2012; Ambawat et al ., 2013; Pattabiraman and Gonda, 2013; Kim et al ., 2014; Baldoni et al ., 2015; Dong et al ., 2015; Liu et al ., 2015; Roy, 2016; Matheis et al ., 2017; Valsecchi et al ., 2017; Verma et al ., 2017; Allan and Espley, 2018; Mitra, 2018; X.…”

“…Wang et al, 2018;Li et al, 2019Li et al, , 2019Ma and Constabel, 2019;Cao et al, 2020) although proteins that are not classical transcription factors can contain Myb type DNA binding domains (Hogues et al, 2008;Yu et al, 2010;Rodríguez-Sánchez et al, 2011;Mukherjee et al, 2012;Pattabiraman and Gonda, 2013;Roy, 2016;X. Wang et al, 2018;Ma and Constabel, 2019;Ramos-Sáenz et al, 2019). The Myb family does not have many members comprising 4 to 5 proteins in animal genomes (Prouse and Campbell, 2012).…”

“…In plants, the Myb family proteins are mainly considered as transcription factors or repressors (Allan and Espley, 2018; Ambawat et al, 2013; Baldoni et al, 2015; Cao et al, 2020; Dong et al, 2015; Du et al, 2009; Dubos et al, 2010; Kim et al, 2014; Li et al, 2019; Li et al, 2019; Lin et al, 2011; Lin et al, 2012; Liu et al, 2015; Ma and Constabel, 2019; Mateos et al, 2006; Matheis et al, 2017; Mitra, 2018; Pattabiraman and Gonda, 2013; Prouse and Campbell, 2012; Roy, 2016; Valsecchi et al, 2017; Verma et al, 2017; X., Wang, et al, 2018; Wieser and Adams, 1995). However, proteins that are not classical transcription factors can contain Myb-type DNA binding domains (Hogues et al, 2008; Ma and Constabel, 2019; Mukherjee et al, 2012; Pattabiraman and Gonda, 2013; Ramos-Sáenz et al, 2019; Rodríguez-Sánchez et al, 2011; Roy, 2016; X., Wang et al, 2018; Yu et al, 2010). The Myb family does not have many members comprising 4 to 5 proteins in animal genomes (Prouse and Campbell, 2012).…”

Non-radioactive In Vivo Labeling of RNA with 4-Thiouracil

“…Please note that depletion of the small ribosomal subunit biogenesis factor, Pwp2, for 14 h leads to the reduction of small ribosomal subunit pre-and mature rRNAs (20S and 18S rRNA, respectively) [39]. In contrast depletion of the large ribosomal subunit biogenesis factor, Pol5, for 9 h leads to the reduction of the common large ribosomal subunit precursor and corresponding mature rRNA (27S, 25S, and 5.8S, respectively) [39][40][41]. Additional application examples using this protocol and S. cerevisiae is described in [41] 1.…”

Bioinformatic Analysis of MYB Transcription Factors in <i>Fusarium graminearum</i>

“…The study of fungal MYB TFs started late, to date they were only reported in the form of single gene in fungi such as yeast, Aspergillus nidulans, Aspergillus fumigatus and Magnaporthe oryzae. Preliminary studies have found that they are essential for fungal spore formation and fungal pathogenicity to plant (Arratia-Quijada et al, 2012;Dong et al, 2018;Sarikaya Bayram et al, 2018;Ramos-Sáenz et al, 2019).…”