Abstract
tRNA 3' processing endoribonuclease (3' tRNase) is an enzyme responsible for the removal of a 3' trailer from precursor tRNA (pre-tRNA). We purified approximately 85 kDa 3' tRNase from pig liver and determined its partial sequences. BLAST search of them suggested that the enzyme was the product of a candidate human prostate cancer susceptibility gene, ELAC2, the biological function of which was totally unknown. We cloned a human ELAC2 cDNA and expressed the ELAC2 protein in Escherichia coli. The recombinant ELAC2 was able to cleave human pre-tRNA(Arg) efficiently. The 3' tRNase activity of the yeast ortholog YKR079C was also observed. The C-terminal half of human ELAC2 was able to remove a 3' trailer from pre-tRNA(Arg), while the N-terminal half failed to do so. In the human genome exists a gene, ELAC1, which seems to correspond to the C-terminal half of 3' tRNase from ELAC2. We showed that human ELAC1 also has 3'-tRNase activity. Furthermore, we examined eight ELAC2 variants that seem to be associated with the occurrence of prostate cancer for 3'-tRNase activity. Seven ELAC2 variants which contain one to three amino acid substitutions showed efficient 3'-tRNase activities, while one truncated variant, which lacked a C-terminal half region, had no activity.
MeSH Terms
Amino Acid Sequence
Animals
Cloning, Molecular
DNA, Complementary/genetics
Endoribonucleases/chemistry,genetics,metabolism
Escherichia coli/genetics
Humans
Male
Molecular Sequence Data
Mutation
Neoplasm Proteins/genetics,metabolism
Nucleic Acid Conformation
Plasmids/genetics
Prostatic Neoplasms/enzymology,genetics
RNA Precursors/chemistry,genetics,metabolism
RNA Processing, Post-Transcriptional
RNA, Transfer, Arg/chemistry,genetics,metabolism
Recombinant Proteins/genetics,isolation & purification,metabolism
Saccharomyces cerevisiae/enzymology,genetics
Sequence Analysis, Protein
Sequence Homology, Amino Acid
Swine
Chemicals
DNA, Complementary
ELAC2 protein, human
Neoplasm Proteins
RNA Precursors
RNA, Transfer, Arg
Recombinant Proteins
Endoribonucleases
3'-pre-tRNase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Takaku Hiroaki
Department of Applied Life Sciences, Niigata University of Pharmacy and Applied Life Sciences, Higashijima 265-1, Niitsu, Niigata 956-8603, Japan.
Minagawa Asako
Takagi Masamichi
Nashimoto Masayuki
References (30)
30 references, click to expand
-
Anomalous RNA substrates for mammalian tRNA 3' processing endoribonuclease.
FEBS Lett. 2000 Apr 28;472(2-3):179-86
PMID: 10788607
-
Selection of cleavage site by mammalian tRNA 3' processing endoribonuclease.
J Mol Biol. 1999 Apr 9;287(4):727-40
PMID: 10191141
-
Cyclic nucleotide phosphodiesterases: relating structure and function.
Prog Nucleic Acid Res Mol Biol. 2001;65:1-52
PMID: 11008484
-
Long 5' leaders inhibit removal of a 3' trailer from a precursor tRNA by mammalian tRNA 3' processing endoribonuclease.
Nucleic Acids Res. 1999 Jul 1;27(13):2770-6
PMID: 10373595
-
Minimum requirements for substrates of mammalian tRNA 3' processing endoribonuclease.
Biochemistry. 1999 Sep 14;38(37):12089-96
PMID: 10508413
-
A candidate prostate cancer susceptibility gene at chromosome 17p.
Nat Genet. 2001 Feb;27(2):172-80
PMID: 11175785
-
Evaluation of linkage and association of HPC2/ELAC2 in patients with familial or sporadic prostate cancer.
Am J Hum Genet. 2001 Apr;68(4):901-11
PMID: 11254448
-
Role of HPC2/ELAC2 in hereditary prostate cancer.
Cancer Res. 2001 Sep 1;61(17):6494-9
PMID: 11522646
-
The inhibitory effect of the autoantigen La on in vitro 3' processing of mammalian precursor tRNAs.
J Mol Biol. 2001 Oct 5;312(5):975-84
PMID: 11580243
-
This is the end: processing, editing and repair at the tRNA 3'-terminus.
Biol Chem. 2001 Aug;382(8):1147-56
PMID: 11592395
-
Assigning a function to a conserved group of proteins: the tRNA 3'-processing enzymes.
EMBO J. 2002 Jun 3;21(11):2769-77
PMID: 12032089
-
Gene silencing in mammals by small interfering RNAs.
Nat Rev Genet. 2002 Oct;3(10):737-47
PMID: 12360232
-
HPC2/ELAC2 polymorphisms and prostate cancer risk: analysis by age of onset of disease.
Br J Cancer. 2002 Oct 7;87(8):905-8
PMID: 12373607
-
Association of common missense changes in ELAC2 ( HPC2) with prostate cancer in a Japanese case-control series.
J Hum Genet. 2002;47(12):641-8
PMID: 12522685
-
The product of the candidate prostate cancer susceptibility gene ELAC2 interacts with the gamma-tubulin complex.
Int J Cancer. 2003 Apr 10;104(3):283-8
PMID: 12569551
-
Direct chemical method for sequencing RNA.
Proc Natl Acad Sci U S A. 1979 Apr;76(4):1760-4
PMID: 377283
-
Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold.
EMBO J. 1982;1(8):945-51
PMID: 6329717
-
Site-directed mutagenesis by overlap extension using the polymerase chain reaction.
Gene. 1989 Apr 15;77(1):51-9
PMID: 2744487
-
A novel spermidine-dependent endoribonuclease activity caused by RNA-protein complex in mouse FM3A cell extracts.
Biochem Biophys Res Commun. 1991 May 15;176(3):1163-9
PMID: 2039502
-
Characterization of the spermidine-dependent, sequence-specific endoribonuclease that requires transfer RNA for its activity.
Nucleic Acids Res. 1992 Jul 25;20(14):3737-42
PMID: 1641339
-
3' truncated tRNAArg is essential for in vitro specific cleavage of partially synthesized mouse 18S rRNA.
Nucleic Acids Res. 1993 Oct 11;21(20):4696-702
PMID: 8233818
-
A single amino acid change in SNM1-encoded protein leads to thermoconditional deficiency for DNA cross-link repair in Saccharomyces cerevisiae.
Mutat Res. 1994 Nov;315(3):275-9
PMID: 7526204
-
Conversion of mammalian tRNA 3' processing endoribonuclease to four-base-recognizing RNA cutters.
Nucleic Acids Res. 1995 Sep 25;23(18):3642-7
PMID: 7478991
-
Specific cleavage of target RNAs from HIV-1 with 5' half tRNA by mammalian tRNA 3' processing endoribonuclease.
RNA. 1996 Jun;2(6):523-4
PMID: 8718682
-
Pre-tRNA 3'-processing in Saccharomyces cerevisiae. Purification and characterization of exo- and endoribonucleases.
Eur J Biochem. 1996 Dec 15;242(3):747-59
PMID: 9022706
-
Distribution of both lengths and 5' terminal nucleotides of mammalian pre-tRNA 3' trailers reflects properties of 3' processing endoribonuclease.
Nucleic Acids Res. 1997 Mar 15;25(6):1148-54
PMID: 9092623
-
The yeast La protein is required for the 3' endonucleolytic cleavage that matures tRNA precursors.
Cell. 1997 May 2;89(3):393-402
PMID: 9150139
-
RNA heptamers that direct RNA cleavage by mammalian tRNA 3' processing endoribonuclease.
Nucleic Acids Res. 1998 Jun 1;26(11):2565-72
PMID: 9592138
-
A zinc-binding motif conserved in glyoxalase II, beta-lactamase and arylsulfatases.
Trends Biochem Sci. 1998 Oct;23(10):381-2
PMID: 9810225
-
Association of HPC2/ELAC2 genotypes and prostate cancer.
Am J Hum Genet. 2000 Oct;67(4):1014-9
PMID: 10986046