Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Science 5 June 1992:
Vol. 256. no. 5062, pp. 1416 - 1419
DOI: 10.1126/science.1604315
Prev | Table of Contents | Next

Articles

Science, Vol 256, Issue 5062, 1416-1419
Copyright © 1992 by American Association for the Advancement of Science

articles

Unusual resistance of peptidyl transferase to protein extraction procedures

HF Noller, V Hoffarth, and L Zimniak

Sinsheimer Laboratories, University of California, Santa Cruz.

Peptidyl transferase, the ribosomal activity responsible for catalysis of peptide bond formation, is resistant to vigorous procedures that are conventionally employed to remove proteins from protein-nucleic acid complexes. When the "fragment reaction" was used as a model assay for peptide bond formation, Escherichia coli ribosomes or 50S subunits retained 20 to 40 percent activity after extensive treatment with proteinase K and SDS, but lost activity after extraction with phenol or exposure to EDTA. Ribosomes from the thermophilic eubacterium Thermus aquaticus remained more than 80 percent active after treatment with proteinase K and SDS, which was followed by vigorous extraction with phenol. This activity is attributable to peptidyl transferase, as judged by specific inhibition by the peptidyl transferase-specific antibiotics chloramphenicol and carbomycin. In contrast, activity is abolished by treatment with ribonuclease T1. These findings support the possibility that 23S ribosomal RNA participates in the peptidyl transferase function.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
The Ribosome: Some Hard Facts about Its Structure and Hot Air about Its Evolution.
V. Ramakrishnan (2009)
Cold Spring Harb Symp Quant Biol
   Abstract »    PDF »
Large facilities and the evolving ribosome, the cellular machine for genetic-code translation.
A. Yonath (2009)
J R Soc Interface 6, S575-S585
   Abstract »    Full Text »    PDF »
The spliceosome as ribozyme hypothesis takes a second step.
S. E. Butcher (2009)
PNAS 106, 12211-12212
   Full Text »    PDF »
A recurrent magnesium-binding motif provides a framework for the ribosomal peptidyl transferase center.
C. Hsiao and L. D. Williams (2009)
Nucleic Acids Res. 37, 3134-3142
   Abstract »    Full Text »    PDF »
Structural effects of nucleobase variations at key active site residue Ade38 in the hairpin ribozyme.
C. MacElrevey, J. D. Salter, J. Krucinska, and J. E. Wedekind (2008)
RNA 14, 1600-1616
   Abstract »    Full Text »    PDF »
Modulating the activity of the peptidyl transferase center of the ribosome.
M. Beringer (2008)
RNA 14, 795-801
   Abstract »    Full Text »    PDF »
Protein Folding by Domain V of Escherichia coli 23S rRNA: Specificity of RNA-Protein Interactions.
D. Samanta, D. Mukhopadhyay, S. Chowdhury, J. Ghosh, S. Pal, A. Basu, A. Bhattacharya, A. Das, D. Das, and C. DasGupta (2008)
J. Bacteriol. 190, 3344-3352
   Abstract »    Full Text »    PDF »
A novel chromatography system to isolate active ribosomes from pathogenic bacteria.
B. A. Maguire, L. M. Wondrack, L. G. Contillo, and Z. Xu (2008)
RNA 14, 188-195
   Abstract »    Full Text »    PDF »
Functional genetic selection of Helix 66 in Escherichia coli 23S rRNA identified the eukaryotic-binding sequence for ribosomal protein L2.
K. Kitahara, A. Kajiura, N. S. Sato, and T. Suzuki (2007)
Nucleic Acids Res. 35, 4018-4029
   Abstract »    Full Text »    PDF »
Evolutionary rates vary among rRNA structural elements.
S. Smit, J. Widmann, and R. Knight (2007)
Nucleic Acids Res. 35, 3339-3354
   Abstract »    Full Text »    PDF »
Comprehensive genetic selection revealed essential bases in the peptidyl-transferase center.
N. S. Sato, N. Hirabayashi, I. Agmon, A. Yonath, and T. Suzuki (2006)
PNAS 103, 15386-15391
   Abstract »    Full Text »    PDF »
Localization of spermine binding sites in 23S rRNA by photoaffinity labeling: parsing the spermine contribution to ribosomal 50S subunit functions.
M. A. Xaplanteri, A. D. Petropoulos, G. P. Dinos, and D. L. Kalpaxis (2005)
Nucleic Acids Res. 33, 2792-2805
   Abstract »    Full Text »    PDF »
The driving force for molecular evolution of translation.
H. F. NOLLER (2004)
RNA 10, 1833-1837
   Abstract »    Full Text »    PDF »
The A2453-C2499 wobble base pair in Escherichia coli 23S ribosomal RNA is responsible for pH sensitivity of the peptidyltransferase active site conformation.
M. A. Bayfield, J. Thompson, and A. E. Dahlberg (2004)
Nucleic Acids Res. 32, 5512-5518
   Abstract »    Full Text »    PDF »
Importance in catalysis of a magnesium ion with very low affinity for a hammerhead ribozyme.
A. Inoue, Y. Takagi, and K. Taira (2004)
Nucleic Acids Res. 32, 4217-4223
   Abstract »    Full Text »    PDF »
Inhibition of Escherichia coli RNase P by oligonucleotide directed misfolding of RNA.
J. L. CHILDS, A. W. POOLE, and D. H. TURNER (2003)
RNA 9, 1437-1445
   Abstract »    Full Text »    PDF »
Evidence against stabilization of the transition state oxyanion by a pKa-perturbed RNA base in the peptidyl transferase center.
K. M. Parnell, A. C. Seila, and S. A. Strobel (2002)
PNAS 99, 11658-11663
   Abstract »    Full Text »    PDF »
Evolutionary Conservation of Reactions in Translation.
M. C. Ganoza, M. C. Kiel, and H. Aoki (2002)
Microbiol. Mol. Biol. Rev. 66, 460-485
   Abstract »    Full Text »    PDF »
Oligonucleotide directed misfolding of RNA inhibits Candida albicans group I intron splicing.
J. L. Childs, M. D. Disney, and D. H. Turner (2002)
PNAS 99, 11091-11096
   Abstract »    Full Text »    PDF »
Existence of efficient divalent metal ion-catalyzed and inefficient divalent metal ion-independent channels in reactions catalyzed by a hammerhead ribozyme.
J.-M. Zhou, D.-M. Zhou, Y. Takagi, Y. Kasai, A. Inoue, T. Baba, and K. Taira (2002)
Nucleic Acids Res. 30, 2374-2382
   Abstract »    Full Text »    PDF »
Minihelix-loop RNAs: minimal structures for aminoacylation catalysts.
K. Ramaswamy, K. Wei, and H. Suga (2002)
Nucleic Acids Res. 30, 2162-2171
   Abstract »    Full Text »    PDF »
Point Mutation in Essential Genes with Loss or Mutation of the Second Allele: Relevance to the Retention of Tumor-Specific Antigens.
G. B. Beck-Engeser, P. A. Monach, D. Mumberg, F. Yang, S. Wanderling, K. Schreiber, R. Espinosa III, M. M. Le Beau, S. C. Meredith, and H. Schreiber (2001)
J. Exp. Med. 194, 285-300
   Abstract »    Full Text »    PDF »
SURVEY AND SUMMARY: Recent advances in the elucidation of the mechanisms of action of ribozymes.
Y. Takagi, M. Warashina, W. J. Stec, K. Yoshinari, and K. Taira (2001)
Nucleic Acids Res. 29, 1815-1834
   Abstract »    Full Text »    PDF »
A Progress Report on Translational Control in Eukaryotes.
M. Kozak (2001)
Sci. STKE 2001, pe1
   Abstract »    Full Text »    PDF »
Oligonucleotide-directed peptide synthesis in a ribosome- and ribozyme-free system.
K. Tamura and P. Schimmel (2001)
PNAS 98, 1393-1397
   Abstract »    Full Text »    PDF »
High-resolution Structures of Ribosomal Subunits: Initiation, Inhibition, and Conformational Variability.
A. BASHAN, I. AGMON, R. ZARIVACH, F. SCHLUENZEN, J. HARMS, M. PIOLETTI, H. BARTELS, M. GLUEHMANN, H. HANSEN, T. AUERBACH, et al. (2001)
Cold Spring Harb Symp Quant Biol 66, 43-56
   Abstract »    PDF »
Probing Ribosome Structure and Function by Mutagenesis.
S.T. GREGORY, M.A. BAYFIELD, M. O'CONNOR, J. THOMPSON, and A.E. DAHLBERG (2001)
Cold Spring Harb Symp Quant Biol 66, 101-108
   Abstract »    PDF »
Codon Recognition and Decoding: The Transorientation Hypothesis.
A.B. SIMONSON and J.A. LAKE (2001)
Cold Spring Harb Symp Quant Biol 66, 127-134
   Abstract »    PDF »
Ribosome Biogenesis: Role of Small Nucleolar RNA in Maturation of Eukaryotic rRNA.
S.A. GERBI, A.V. BOROVJAGIN, M. EZROKHI, and T.S. LANGE (2001)
Cold Spring Harb Symp Quant Biol 66, 575-590
   Abstract »    PDF »
The Structural Basis of Ribosome Activity in Peptide Bond Synthesis.
P. Nissen, J. Hansen, N. Ban, P. B. Moore, and T. A. Steitz (2000)
Science 289, 920-930
   Abstract »    Full Text »
Complementary Role of Two Fragments of Domain V of 23 S Ribosomal RNA in Protein Folding.
S. Pal, S. Chandra, S. Chowdhury, D. Sarkar, A. N. Ghosh, and C. D. Gupta (1999)
J. Biol. Chem. 274, 32771-32777
   Abstract »    Full Text »    PDF »
Specific, rapid synthesis of Phe-RNA by RNA.
M. Illangasekare and M. Yarus (1999)
PNAS 96, 5470-5475
   Abstract »    Full Text »    PDF »
Engineering of bacterial ribosomes: Replacement of all seven Escherichia coli rRNA operons by a single plasmid-encoded operon.
M. Nomura (1999)
PNAS 96, 1820-1822
   Full Text »    PDF »
An Escherichia coli strain with all chromosomal rRNA operons inactivated: Complete exchange of rRNA genes between bacteria.
T. Asai, D. Zaporojets, C. Squires, and C. L. Squires (1999)
PNAS 96, 1971-1976
   Abstract »    Full Text »    PDF »
Genetic code origins: Experiments confirm phylogenetic predictions and may explain a puzzle.
P. Schimmel and L. Ribas de Pouplana (1999)
PNAS 96, 327-328
   Full Text »    PDF »
Characterization of functionally active subribosomal particles from Thermus aquaticus.
P. Khaitovich, A. S. Mankin, R. Green, L. Lancaster, and H. F. Noller (1999)
PNAS 96, 85-90
   Abstract »    Full Text »    PDF »
Genetic code origins: tRNAs older than their synthetases?.
L. R. de Pouplana, R. J. Turner, B. A. Steer, and P. Schimmel (1998)
PNAS 95, 11295-11300
   Abstract »    Full Text »    PDF »
Reconstitution of Peptide Bond Formation with Escherichia coli 23S Ribosomal RNA Domains.
I. Nitta, Y. Kamada, H. Noda, T. Ueda, and K. Watanabe (1998)
Science 281, 666-669
   Abstract »    Full Text »
Mass spectrometry of ribosomes and ribosomal subunits.
D. R. Benjamin, C. V. Robinson, J. P. Hendrick, F. U. Hartl, and C. M. Dobson (1998)
PNAS 95, 7391-7395
   Abstract »    Full Text »    PDF »
An Intragenic Suppressor of Cold Sensitivity Identifies Potentially Interacting Bases in the Peptidyl Transferase Center of Tetrahymena rRNA.
R. Sweeney and M.-C. Yao (1998)
Genetics 149, 937-946
   Abstract »    Full Text »    PDF »
An rRNA Fragment and Its Antisense Can Alter Decoding of Genetic Information.
A. L. Arkov, A. Mankin, and E. J. Murgola (1998)
J. Bacteriol. 180, 2744-2748
   Abstract »    Full Text »
Ribosome-Catalyzed Peptide-Bond Formation with an A-Site Substrate Covalently Linked to 23S Ribosomal RNA.
R. Green, C. Switzer, and H. F. Noller (1998)
Science 280, 286-289
   Abstract »    Full Text »
Inhibition of translation and bacterial growth by peptide nucleic acid targeted to ribosomal RNA.
L. Good and P. E. Nielsen (1998)
PNAS 95, 2073-2076
   Abstract »    Full Text »    PDF »
The Gene Encoding the Elongation Factor P Protein Is Essential for Viability and Is Required for Protein Synthesis.
H. Aoki, K. Dekany, S.-L. Adams, and M. C. Ganoza (1997)
J. Biol. Chem. 272, 32254-32259
   Abstract »    Full Text »    PDF »
In vitro selection of self-cleaving RNAs with a low pH optimum.
V. K. Jayasena and L. Gold (1997)
PNAS 94, 10612-10617
   Abstract »    Full Text »    PDF »
A protein encoded by a group I intron in Aspergillus nidulans directly assists RNA splicing and is a DNA endonuclease.
Y. Ho, S.-J. Kim, and R. B. Waring (1997)
PNAS 94, 8994-8999
   Abstract »    Full Text »    PDF »
Cytokinin Induction of RNA Polymerase I Transcription in Arabidopsis thaliana.
R. J. Gaudino and C. S. Pikaard (1997)
J. Biol. Chem. 272, 6799-6804
   Abstract »    Full Text »    PDF »
Mutational Analysis of Two Highly Conserved UGG Sequences of 23S rRNA from Escherichia coli.
C. M.T. Spahn, J. Remme, M. A. Schafer, and KnudH. Nierhaus (1996)
J. Biol. Chem. 271, 32849-32856
   Abstract »    Full Text »    PDF »
The Leucyl/Phenylalanyl-tRNA-Protein Transferase.
G. Abramochkin and T. E. Shrader (1995)
J. Biol. Chem. 270, 20621-20628
   Abstract »    Full Text »    PDF »
Posttranscriptional Modification of the Central Loop of Domain V in Escherichia coli 23 S Ribosomal RNA.
J. A. Kowalak, E. Bruenger, and J. A. McCloskey (1995)
J. Biol. Chem. 270, 17758-17764
   Abstract »    Full Text »    PDF »
Aminoacyl-RNA synthesis catalyzed by an RNA.
M Illangasekare, G Sanchez, T Nickles, and M Yarus (1995)
Science 267, 643-647
   Abstract »    PDF »
Cleavage of an amide bond by a ribozyme.
X Dai, A De Mesmaeker, and G. Joyce (1995)
Science 267, 237-240
   Abstract »    PDF »
Expanding the scope of RNA catalysis.
Prudent JR, T Uno, and P. Schultz (1994)
Science 264, 1924-1927
   Abstract »    PDF »
Functional reconstitution of wild-type and mutant Tetrahymena telomerase..
C Autexier and C W Greider (1994)
Genes & Dev. 8, 563-575
   Abstract »    PDF »
Functional requirement of a site-specific ribose methylation in ribosomal RNA.
K Sirum-Connolly and T. Mason (1993)
Science 262, 1886-1889
   Abstract »    PDF »
Ribozymes: a distinct class of metalloenzymes.
A. Pyle (1993)
Science 261, 709-714
   Abstract »    PDF »
From RNA to DNA, why so many ribonucleotide reductases?.
P Reichard (1993)
Science 260, 1773-1777
   Abstract »    PDF »
Splice-site selection and decoding: are they related?.
R Schroeder, B Streicher, and H Wank (1993)
Science 260, 1443-1444
   PDF »
Movement of the guide sequence during RNA catalysis by a group I ribozyme.
J. Wang, W. Downs, and T. Cech (1993)
Science 260, 504-508
   Abstract »    PDF »
An enhancer screen identifies a gene that encodes the yeast U1 snRNP A protein: implications for snRNP protein function in pre-mRNA splicing..
X C Liao, J Tang, and M Rosbash (1993)
Genes & Dev. 7, 419-428
   Abstract »    PDF »
Biologists trace the evolution of molecules.
A Gibbons (1992)
Science 257, 30-31
   PDF »
New horizons for RNA catalysis.
N. Pace (1992)
Science 256, 1402-1403
   PDF »
Aminoacyl esterase activity of the Tetrahymena ribozyme.
J. Piccirilli, T. McConnell, A. Zaug, H. Noller, and T. Cech (1992)
Science 256, 1420-1424
   Abstract »    PDF »
Crystal Structure of the Ribosome at 5.5 A Resolution.
M. M. Yusupov, G. Zh. Yusupova, A. Baucom, K. Lieberman, T. N. Earnest, J. H. D. Cate, and H. F. Noller (2001)
Science 292, 883-896
   Abstract »    Full Text »    PDF »
Functional Equivalence of Structurally Distinct Ribosomes in the Malaria Parasite, Plasmodium berghei.
R. M. L. van Spaendonk, J. Ramesar, A. van Wigcheren, W. Eling, A. L. Beetsma, G.-J. van Gemert, J. Hooghof, C. J. Janse, and A. P. Waters (2001)
J. Biol. Chem. 276, 22638-22647
   Abstract »    Full Text »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)