Recognition determinants for proteins and antibiotics within 23S rRNA

Stephen Roger Douthwaite, Bjørn Gunnar Rude Voldborg, Lykke Haastrup Hansen, G Rosendahl, B Vester

Publikation: Bidrag til tidsskriftKonferenceartikelForskningpeer review

Resumé

Ribosomal RNAs fold into phylogenetically conserved secondary and tertiary structures that determine their function in protein synthesis. We have investigated Escherichia coli 23S rRNA to identify structural elements that interact with antibiotic and protein ligands. Using a combination of molecular genetic and biochemical probing techniques, we have concentrated on regions of the rRNA that are connected with specific functions. These are located in different domains within the 23S rRNA and include the ribosomal GTPase-associated center in domain II, which contains the binding sites for r-proteins L10.(L12)4 and L11 and is inhibited by interaction with the antibiotic thiostrepton. The peptidyltransferase center within domain V is inhibited by macrolide, lincosamide, and streptogramin B antibiotics, which interact with the rRNA around nucleotide A2058. Drug resistance is conferred by mutations here and by modification of A2058 by ErmE methyltransferase. ErmE recognizes a conserved motif displayed in the primary and secondary structure of the peptidyl transferase loop. Within domain VI of rRNA, the alpha-sarcin stem-loop is associated with elongation factor binding and is the target site for ribotoxins including the N-glycosidase ribosome-inactivating proteins ricin and pokeweed antiviral protein (PAP). The orientations of the 23S rRNA domains are constrained by tetiary interactions, including a pseudoknot in domain II and long-range base pairings in the center of the molecule that bring domains II and V closer together. The phenotypic effects of mutations in these regions have been investigated by expressing 23S rRNA from plasmids. Allele-specific priming sites have been introduced close to these structures in the rRNA to enable us to study the molecular events there.
OriginalsprogEngelsk
TidsskriftBiochemistry and Cell Biology
Vol/bind73
Udgave nummer11-12
Sider (fra-til)1179-85
Antal sider7
ISSN0829-8211
StatusUdgivet - 1995

Fingeraftryk

Peptidyl Transferases
Anti-Bacterial Agents
Thiostrepton
Streptogramin B
Lincosamides
Ribosome Inactivating Proteins
Peptide Elongation Factors
Ricin
Mutation
Proteins
Ribosomal RNA
Glycoside Hydrolases
GTP Phosphohydrolases
Macrolides
Drug Resistance
Base Pairing
Escherichia coli
Plasmids
Nucleotides
Alleles

Citer dette

Douthwaite, Stephen Roger ; Voldborg, Bjørn Gunnar Rude ; Hansen, Lykke Haastrup ; Rosendahl, G ; Vester, B. / Recognition determinants for proteins and antibiotics within 23S rRNA. I: Biochemistry and Cell Biology. 1995 ; Bind 73, Nr. 11-12. s. 1179-85.
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title = "Recognition determinants for proteins and antibiotics within 23S rRNA",
abstract = "Ribosomal RNAs fold into phylogenetically conserved secondary and tertiary structures that determine their function in protein synthesis. We have investigated Escherichia coli 23S rRNA to identify structural elements that interact with antibiotic and protein ligands. Using a combination of molecular genetic and biochemical probing techniques, we have concentrated on regions of the rRNA that are connected with specific functions. These are located in different domains within the 23S rRNA and include the ribosomal GTPase-associated center in domain II, which contains the binding sites for r-proteins L10.(L12)4 and L11 and is inhibited by interaction with the antibiotic thiostrepton. The peptidyltransferase center within domain V is inhibited by macrolide, lincosamide, and streptogramin B antibiotics, which interact with the rRNA around nucleotide A2058. Drug resistance is conferred by mutations here and by modification of A2058 by ErmE methyltransferase. ErmE recognizes a conserved motif displayed in the primary and secondary structure of the peptidyl transferase loop. Within domain VI of rRNA, the alpha-sarcin stem-loop is associated with elongation factor binding and is the target site for ribotoxins including the N-glycosidase ribosome-inactivating proteins ricin and pokeweed antiviral protein (PAP). The orientations of the 23S rRNA domains are constrained by tetiary interactions, including a pseudoknot in domain II and long-range base pairings in the center of the molecule that bring domains II and V closer together. The phenotypic effects of mutations in these regions have been investigated by expressing 23S rRNA from plasmids. Allele-specific priming sites have been introduced close to these structures in the rRNA to enable us to study the molecular events there.",
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year = "1995",
language = "English",
volume = "73",
pages = "1179--85",
journal = "Biochemistry and Cell Biology",
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Douthwaite, SR, Voldborg, BGR, Hansen, LH, Rosendahl, G & Vester, B 1995, 'Recognition determinants for proteins and antibiotics within 23S rRNA', Biochemistry and Cell Biology, bind 73, nr. 11-12, s. 1179-85.

Recognition determinants for proteins and antibiotics within 23S rRNA. / Douthwaite, Stephen Roger; Voldborg, Bjørn Gunnar Rude; Hansen, Lykke Haastrup; Rosendahl, G; Vester, B.

I: Biochemistry and Cell Biology, Bind 73, Nr. 11-12, 1995, s. 1179-85.

Publikation: Bidrag til tidsskriftKonferenceartikelForskningpeer review

TY - GEN

T1 - Recognition determinants for proteins and antibiotics within 23S rRNA

AU - Douthwaite, Stephen Roger

AU - Voldborg, Bjørn Gunnar Rude

AU - Hansen, Lykke Haastrup

AU - Rosendahl, G

AU - Vester, B

PY - 1995

Y1 - 1995

N2 - Ribosomal RNAs fold into phylogenetically conserved secondary and tertiary structures that determine their function in protein synthesis. We have investigated Escherichia coli 23S rRNA to identify structural elements that interact with antibiotic and protein ligands. Using a combination of molecular genetic and biochemical probing techniques, we have concentrated on regions of the rRNA that are connected with specific functions. These are located in different domains within the 23S rRNA and include the ribosomal GTPase-associated center in domain II, which contains the binding sites for r-proteins L10.(L12)4 and L11 and is inhibited by interaction with the antibiotic thiostrepton. The peptidyltransferase center within domain V is inhibited by macrolide, lincosamide, and streptogramin B antibiotics, which interact with the rRNA around nucleotide A2058. Drug resistance is conferred by mutations here and by modification of A2058 by ErmE methyltransferase. ErmE recognizes a conserved motif displayed in the primary and secondary structure of the peptidyl transferase loop. Within domain VI of rRNA, the alpha-sarcin stem-loop is associated with elongation factor binding and is the target site for ribotoxins including the N-glycosidase ribosome-inactivating proteins ricin and pokeweed antiviral protein (PAP). The orientations of the 23S rRNA domains are constrained by tetiary interactions, including a pseudoknot in domain II and long-range base pairings in the center of the molecule that bring domains II and V closer together. The phenotypic effects of mutations in these regions have been investigated by expressing 23S rRNA from plasmids. Allele-specific priming sites have been introduced close to these structures in the rRNA to enable us to study the molecular events there.

AB - Ribosomal RNAs fold into phylogenetically conserved secondary and tertiary structures that determine their function in protein synthesis. We have investigated Escherichia coli 23S rRNA to identify structural elements that interact with antibiotic and protein ligands. Using a combination of molecular genetic and biochemical probing techniques, we have concentrated on regions of the rRNA that are connected with specific functions. These are located in different domains within the 23S rRNA and include the ribosomal GTPase-associated center in domain II, which contains the binding sites for r-proteins L10.(L12)4 and L11 and is inhibited by interaction with the antibiotic thiostrepton. The peptidyltransferase center within domain V is inhibited by macrolide, lincosamide, and streptogramin B antibiotics, which interact with the rRNA around nucleotide A2058. Drug resistance is conferred by mutations here and by modification of A2058 by ErmE methyltransferase. ErmE recognizes a conserved motif displayed in the primary and secondary structure of the peptidyl transferase loop. Within domain VI of rRNA, the alpha-sarcin stem-loop is associated with elongation factor binding and is the target site for ribotoxins including the N-glycosidase ribosome-inactivating proteins ricin and pokeweed antiviral protein (PAP). The orientations of the 23S rRNA domains are constrained by tetiary interactions, including a pseudoknot in domain II and long-range base pairings in the center of the molecule that bring domains II and V closer together. The phenotypic effects of mutations in these regions have been investigated by expressing 23S rRNA from plasmids. Allele-specific priming sites have been introduced close to these structures in the rRNA to enable us to study the molecular events there.

KW - Anti-Bacterial Agents

KW - Base Sequence

KW - Binding Sites

KW - Escherichia coli

KW - Molecular Sequence Data

KW - Nucleic Acid Conformation

KW - RNA, Bacterial

KW - RNA, Ribosomal, 23S

KW - RNA-Binding Proteins

M3 - Conference article

VL - 73

SP - 1179

EP - 1185

JO - Biochemistry and Cell Biology

JF - Biochemistry and Cell Biology

SN - 0829-8211

IS - 11-12

ER -