TY - JOUR
T1 - TbUTP10, a protein involved in early stages of pre-18S rRNA processing in Trypanosoma brucei
AU - Faktorová, Drahomíra
AU - Bär, Anita
AU - Hashimi, Hassan
AU - McKenney, Katherine
AU - Horák, Aleš
AU - Schnaufer, Achim
AU - Rubio, Mary Anne T
AU - Alfonzo, Juan D
AU - Lukeš, Julius
N1 - All rights reserved - Publisher requires the AAM to be subject to a 12 month embargo and at the point of release apply the CC-BY-NC-ND licence - Contact Researcher to ask reason for late deposit 18/04/2019 EN - email sent 3/05/2019 EN - Researcher forgot about letting us know 6/05/2019 EN - If the Deposit date has been recorded in PubMed it could be compliant 6/05/2019 EN
PY - 2018/10/1
Y1 - 2018/10/1
N2 - Ribosome biosynthesis, best studied in opisthokonts, is a highly complex process involving numerous protein and RNA factors. Yet, very little is known about the early stages of pre-18S rRNA processing even in these model organisms, let alone the conservation of this mechanism in other eukaryotes. Here we extend our knowledge of this process by identifying and characterizing the essential protein TbUTP10, a homolog of yeast U3 small nucleolar RNA-associated protein 10 - UTP10 (HEATR1 in human), in the excavate parasitic protist Trypanosoma brucei. We show that TbUTP10 localizes to the nucleolus and that its ablation by RNAi knock-down in two different T. brucei life cycle stages results in similar phenotypes: a disruption of pre-18S rRNA processing, exemplified by the accumulation of rRNA precursors, a reduction of mature 18S rRNA, and also a decrease in the level of U3 snoRNA. Moreover, polysome profiles of the RNAi-induced knock-down cells show a complete disappearance of the 40S ribosomal subunit, and a prominent accumulation of the 60S large ribosomal subunit, reflecting impaired ribosome assembly. Thus, TbUTP10 is an important protein in the processing of 18S rRNA.
AB - Ribosome biosynthesis, best studied in opisthokonts, is a highly complex process involving numerous protein and RNA factors. Yet, very little is known about the early stages of pre-18S rRNA processing even in these model organisms, let alone the conservation of this mechanism in other eukaryotes. Here we extend our knowledge of this process by identifying and characterizing the essential protein TbUTP10, a homolog of yeast U3 small nucleolar RNA-associated protein 10 - UTP10 (HEATR1 in human), in the excavate parasitic protist Trypanosoma brucei. We show that TbUTP10 localizes to the nucleolus and that its ablation by RNAi knock-down in two different T. brucei life cycle stages results in similar phenotypes: a disruption of pre-18S rRNA processing, exemplified by the accumulation of rRNA precursors, a reduction of mature 18S rRNA, and also a decrease in the level of U3 snoRNA. Moreover, polysome profiles of the RNAi-induced knock-down cells show a complete disappearance of the 40S ribosomal subunit, and a prominent accumulation of the 60S large ribosomal subunit, reflecting impaired ribosome assembly. Thus, TbUTP10 is an important protein in the processing of 18S rRNA.
KW - Gene Silencing
KW - Genes, Essential
KW - Protozoan Proteins/genetics
KW - RNA Processing, Post-Transcriptional
KW - RNA, Ribosomal, 18S/metabolism
KW - RNA, Small Nucleolar/metabolism
KW - RNA-Binding Proteins/genetics
KW - Trypanosoma brucei brucei/enzymology
U2 - 10.1016/j.molbiopara.2018.09.003
DO - 10.1016/j.molbiopara.2018.09.003
M3 - Article
C2 - 30248370
SN - 0166-6851
VL - 225
SP - 84
EP - 93
JO - Molecular and Biochemical Parasitology
JF - Molecular and Biochemical Parasitology
ER -