Dbp10p, a Putative RNA Helicase from Saccharomyces Cerevisiae, is Required for Ribosome Biogenesis

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2000 Jun 28

PMID 10871363

Citations 31

Authors

F Burger

M C Daugeron

P Linder

Affiliations

Soon will be listed here.

Abstract

Ribosome biogenesis requires, in addition to rRNA molecules and ribosomal proteins, a multitude of trans-acting factors. Recently it has become clear that in the yeast Saccharomyces cerevisiae many RNA helicases of the DEAD-box and related families are involved in ribosome biogenesis. Here we show that the previously uncharacterised open reading frame YDL031w (renamed DBP10 for DEAD-box protein 10) encodes an essential putative RNA helicase that is required for accurate ribosome biogenesis. Genetic depletion of Dbp10p results in a deficit in 60S ribosomal subunits and an accumulation of half-mer polysomes. Furthermore, pulse-chase analyses of pre-rRNA processing reveal a strong delay in the maturation of 27SB pre-rRNA intermediates into 25S rRNA and 7S pre-rRNA. Northern blot analyses indicate that this delay leads to higher steady-state levels of 27SB species and reduced steady-state levels of 7S pre-rRNA and 25S/5.8S mature rRNAs, thus explaining the final deficit in 60S subunit and the formation of half-mer polysomes. Consistent with a direct role in ribosome biogenesis, Dbp10p was found to be located predominantly in the nucleolus.

Citing Articles

Tracing the evolution and genomic dynamics of mating-type loci in pathogens and closely related species.

Coelho M, David-Palma M, Marincowitz S, Aylward J, Pham N, Yurkov A bioRxiv. 2025; .

PMID: 39990455 PMC: 11844451. DOI: 10.1101/2025.02.12.637874.

The RNA helicase Dbp10 coordinates assembly factor association with PTC maturation during ribosome biogenesis.

Mitterer V, Hamze H, Kunowska N, Stelzl U, Henras A, Hurt E Nucleic Acids Res. 2023; 52(4):1975-1987.

PMID: 38113283 PMC: 10899779. DOI: 10.1093/nar/gkad1206.

Nucleolar dynamics are determined by the ordered assembly of the ribosome.

Sheu-Gruttadauria J, Yan X, Stuurman N, Vale R, Floor S bioRxiv. 2023; .

PMID: 37808656 PMC: 10557630. DOI: 10.1101/2023.09.26.559432.

Mechanism of 5S RNP recruitment and helicase-surveilled rRNA maturation during pre-60S biogenesis.

Lau B, Huang Z, Kellner N, Niu S, Berninghausen O, Beckmann R EMBO Rep. 2023; 24(7):e56910.

PMID: 37129998 PMC: 10328080. DOI: 10.15252/embr.202356910.

Sequence-specific remodeling of a topologically complex RNP substrate by Spb4.

Cruz V, Sekulski K, Peddada N, Sailer C, Balasubramanian S, Weirich C Nat Struct Mol Biol. 2022; 29(12):1228-1238.

PMID: 36482249 PMC: 10680166. DOI: 10.1038/s41594-022-00874-9.

References

Trapman J, Planta R, Raue H . Maturation of ribosomes in yeast. II. Position of the low molecular weight rRNA species in the maturation process. Biochim Biophys Acta. 1976; 442(3):275-84. DOI: 10.1016/0005-2787(76)90302-6. View

Li B, Nierras C, Warner J . Transcriptional elements involved in the repression of ribosomal protein synthesis. Mol Cell Biol. 1999; 19(8):5393-404. PMC: 84382. DOI: 10.1128/MCB.19.8.5393. View

Thomas B, Rothstein R . Elevated recombination rates in transcriptionally active DNA. Cell. 1989; 56(4):619-30. DOI: 10.1016/0092-8674(89)90584-9. View

Moritz M, Paulovich A, Tsay Y, Woolford Jr J . Depletion of yeast ribosomal proteins L16 or rp59 disrupts ribosome assembly. J Cell Biol. 1990; 111(6 Pt 1):2261-74. PMC: 2116383. DOI: 10.1083/jcb.111.6.2261. View

Warner J . The economics of ribosome biosynthesis in yeast. Trends Biochem Sci. 1999; 24(11):437-40. DOI: 10.1016/s0968-0004(99)01460-7. View

Kressler D, Rojo M, Linder P, Cruz J . Spb1p is a putative methyltransferase required for 60S ribosomal subunit biogenesis in Saccharomyces cerevisiae. Nucleic Acids Res. 1999; 27(23):4598-608. PMC: 148748. DOI: 10.1093/nar/27.23.4598. View

Kressler D, Linder P, de la Cruz J . Protein trans-acting factors involved in ribosome biogenesis in Saccharomyces cerevisiae. Mol Cell Biol. 1999; 19(12):7897-912. PMC: 84875. DOI: 10.1128/MCB.19.12.7897. View

Kressler D, Doere M, Rojo M, Linder P . Synthetic lethality with conditional dbp6 alleles identifies rsa1p, a nucleoplasmic protein involved in the assembly of 60S ribosomal subunits. Mol Cell Biol. 1999; 19(12):8633-45. PMC: 85000. DOI: 10.1128/MCB.19.12.8633. View

Pintard L, Kressler D, Lapeyre B . Spb1p is a yeast nucleolar protein associated with Nop1p and Nop58p that is able to bind S-adenosyl-L-methionine in vitro. Mol Cell Biol. 2000; 20(4):1370-81. PMC: 85287. DOI: 10.1128/MCB.20.4.1370-1381.2000. View

10.

Venema J, Tollervey D . Ribosome synthesis in Saccharomyces cerevisiae. Annu Rev Genet. 2000; 33:261-311. DOI: 10.1146/annurev.genet.33.1.261. View

11.

Hirling H, Scheffner M, Restle T, Stahl H . RNA helicase activity associated with the human p68 protein. Nature. 1989; 339(6225):562-4. DOI: 10.1038/339562a0. View

12.

Ho S, Hunt H, Horton R, Pullen J, Pease L . Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene. 1989; 77(1):51-9. DOI: 10.1016/0378-1119(89)90358-2. View

13.

Robbins J, Dilworth S, Laskey R, Dingwall C . Two interdependent basic domains in nucleoplasmin nuclear targeting sequence: identification of a class of bipartite nuclear targeting sequence. Cell. 1991; 64(3):615-23. DOI: 10.1016/0092-8674(91)90245-t. View

14.

Gietz D, St Jean A, Woods R, Schiestl R . Improved method for high efficiency transformation of intact yeast cells. Nucleic Acids Res. 1992; 20(6):1425. PMC: 312198. DOI: 10.1093/nar/20.6.1425. View

15.

Ripmaster T, Vaughn G, Woolford Jr J . A putative ATP-dependent RNA helicase involved in Saccharomyces cerevisiae ribosome assembly. Proc Natl Acad Sci U S A. 1992; 89(23):11131-5. PMC: 50503. DOI: 10.1073/pnas.89.23.11131. View

16.

Deshmukh M, Tsay Y, Paulovich A, Woolford Jr J . Yeast ribosomal protein L1 is required for the stability of newly synthesized 5S rRNA and the assembly of 60S ribosomal subunits. Mol Cell Biol. 1993; 13(5):2835-45. PMC: 359670. DOI: 10.1128/mcb.13.5.2835-2845.1993. View

17.

Liang L, Diehl-Jones W, Lasko P . Localization of vasa protein to the Drosophila pole plasm is independent of its RNA-binding and helicase activities. Development. 1994; 120(5):1201-11. DOI: 10.1242/dev.120.5.1201. View

18.

Berges T, Petfalski E, Tollervey D, Hurt E . Synthetic lethality with fibrillarin identifies NOP77p, a nucleolar protein required for pre-rRNA processing and modification. EMBO J. 1994; 13(13):3136-48. PMC: 395205. DOI: 10.1002/j.1460-2075.1994.tb06612.x. View

19.

Ohtake Y, Wickner R . Yeast virus propagation depends critically on free 60S ribosomal subunit concentration. Mol Cell Biol. 1995; 15(5):2772-81. PMC: 230508. DOI: 10.1128/MCB.15.5.2772. View

20.

Wach A, Brachat A, Pohlmann R, Philippsen P . New heterologous modules for classical or PCR-based gene disruptions in Saccharomyces cerevisiae. Yeast. 1994; 10(13):1793-808. DOI: 10.1002/yea.320101310. View