Interactions among Ytm1, Erb1, and Nop7 required for assembly of the Nop7-subcomplex in yeast preribosomes.
In Saccharomyces cerevisiae, more than 180 assembly factors associate with preribosomes to enable folding of pre-rRNA, recruitment of ribosomal proteins, and processing of pre-rRNAs to produce mature ribosomes. To examine the molecular architecture of preribosomes and to connect this structure to functions of each assembly factor, assembly subcomplexes have been purified from preribosomal particles. The Nop7-subcomplex contains three assembly factors: Nop7, Erb1, and Ytm1, each of which is necessary for conversion of 27SA(3) pre-rRNA to 27SB(S) pre-rRNA. However, interactions among these three proteins and mechanisms of their recruitment and function in pre-rRNPs are poorly understood. Here we show that Ytm1, Erb1, and Nop7 assemble into preribosomes in an interdependent manner. We identified which domains within Ytm1, Erb1, and Nop7 are necessary for their interaction with each other and are sufficient for recruitment of each protein into preribosomes. Dominant negative effects on growth and ribosome biogenesis caused by overexpressing truncated Ytm1, Erb1, or Nop7 constructs, and recessive phenotypes of the truncated proteins revealed not only interaction domains but also other domains potentially important for each protein to function in ribosome biogenesis. Our data suggest a model for the architecture of the Nop7-subcomplex and provide potential functions of domains of each protein.
Tang
Lan
L
Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, PA 15213, USA.
Sahasranaman
Aarti
A
Jakovljevic
Jelena
J
Schleifman
Erica
E
Woolford
John L
JL
D020816:Amino Acid Motifs; D003001:Cloning, Molecular; D019084:Fluorescent Antibody Technique, Indirect; D015966:Gene Expression Regulation, Fungal; D005799:Genes, Dominant; D008869:Microtubule-Associated Proteins / Q000502:physiology*; D008954:Models, Biological; D008968:Molecular Conformation; D009154:Mutation; D009687:Nuclear Proteins / Q000502:physiology*; D017434:Protein Structure, Tertiary; D012269:Ribosomal Proteins / Q000502:physiology*; D012270:Ribosomes / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000502:physiology*; D013696:Temperature
C506467:Erb1 protein, S cerevisiae; D008869:Microtubule-Associated Proteins; C451799:NOP7 protein, S cerevisiae; D009687:Nuclear Proteins; D012269:Ribosomal Proteins; D029701:Saccharomyces cerevisiae Proteins; C506466:Ytm1 protein, S cerevisiae