Nucleic acid and protein factors involved in Escherichia coli polynucleotide phosphorylase function on RNA. 2010

Fernando Fernández-Ramírez, and Rosa M Bermúdez-Cruz, and Cecilia Montañez
Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN, DF, Mexico.

It has been reported that polynucleotide phosphorylase (PNPase) binds to RNA via KH and S1 domains, and at least two main complexes (I and II) have been observed in RNA-binding assays. Here we describe PNPase binding to RNA, the factors involved in this activity and the nature of the interactions observed in vitro. Our results show that RNA length and composition affect PNPase binding, and that PNPase interacts primarily with the 3' end of RNA, forming the complex I-RNA, which contains trimeric units of PNPase. When the 5' end of RNA is blocked by a hybridizing oligonucleotide, the formation of complex II-RNA is inhibited. In addition, PNPase was found to form high molecular weight (>440 kDa) aggregates in vitro in the absence of RNA, which may correspond to the hexameric form of the enzyme. We confirmed that PNPase in vitro RNA binding, degradation and polyadenylation activities depend on the integrity of KH and S1 domains. These results can explain the defective in vivo autoregulation of PNPase71, a KH point substitution mutant. As previously reported, optimal growth of a cold-sensitive strain at 18 degrees C requires a fully active PNPase, however, we show that overexpression of a novel PNPaseDeltaS1 partially compensated the growth impairment of this strain, while PNPase71 showed a minor compensation effect. Finally, we propose a mechanism of PNPase interactions and discuss their implications in PNPase function.

UI MeSH Term Description Entries
D008970 Molecular Weight The sum of the weight of all the atoms in a molecule. Molecular Weights,Weight, Molecular,Weights, Molecular
D011117 Polyribonucleotide Nucleotidyltransferase An enzyme of the transferase class that catalyzes the reaction RNA(n+1) and orthophosphate to yield RNA(n) and a nucleoside diphosphate, or the reverse reaction. ADP, IDP, GDP, UDP, and CDP can act as donors in the latter case. (From Dorland, 27th ed) EC 2.7.7.8. Polynucleotide Phosphorylase,Nucleotidyltransferase, Polyribonucleotide,Phosphorylase, Polynucleotide
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D001483 Base Sequence The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence. DNA Sequence,Nucleotide Sequence,RNA Sequence,DNA Sequences,Base Sequences,Nucleotide Sequences,RNA Sequences,Sequence, Base,Sequence, DNA,Sequence, Nucleotide,Sequence, RNA,Sequences, Base,Sequences, DNA,Sequences, Nucleotide,Sequences, RNA
D012329 RNA, Bacterial Ribonucleic acid in bacteria having regulatory and catalytic roles as well as involvement in protein synthesis. Bacterial RNA
D015152 Blotting, Northern Detection of RNA that has been electrophoretically separated and immobilized by blotting on nitrocellulose or other type of paper or nylon membrane followed by hybridization with labeled NUCLEIC ACID PROBES. Northern Blotting,Blot, Northern,Northern Blot,Blots, Northern,Blottings, Northern,Northern Blots,Northern Blottings
D015153 Blotting, Western Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes. Immunoblotting, Western,Western Blotting,Western Immunoblotting,Blot, Western,Immunoblot, Western,Western Blot,Western Immunoblot,Blots, Western,Blottings, Western,Immunoblots, Western,Immunoblottings, Western,Western Blots,Western Blottings,Western Immunoblots,Western Immunoblottings
D017931 DNA Primers Short sequences (generally about 10 base pairs) of DNA that are complementary to sequences of messenger RNA and allow reverse transcriptases to start copying the adjacent sequences of mRNA. Primers are used extensively in genetic and molecular biology techniques. DNA Primer,Oligodeoxyribonucleotide Primer,Oligodeoxyribonucleotide Primers,Oligonucleotide Primer,Oligonucleotide Primers,Primer, DNA,Primer, Oligodeoxyribonucleotide,Primer, Oligonucleotide,Primers, DNA,Primers, Oligodeoxyribonucleotide,Primers, Oligonucleotide

Related Publications

Fernando Fernández-Ramírez, and Rosa M Bermúdez-Cruz, and Cecilia Montañez
August 1987, Molecular & general genetics : MGG,
Fernando Fernández-Ramírez, and Rosa M Bermúdez-Cruz, and Cecilia Montañez
June 2011, RNA (New York, N.Y.),
Fernando Fernández-Ramírez, and Rosa M Bermúdez-Cruz, and Cecilia Montañez
November 1963, Biochimica et biophysica acta,
Fernando Fernández-Ramírez, and Rosa M Bermúdez-Cruz, and Cecilia Montañez
July 1964, Biochimica et biophysica acta,
Fernando Fernández-Ramírez, and Rosa M Bermúdez-Cruz, and Cecilia Montañez
November 1961, The Biochemical journal,
Fernando Fernández-Ramírez, and Rosa M Bermúdez-Cruz, and Cecilia Montañez
August 1971, Comptes rendus hebdomadaires des seances de l'Academie des sciences. Serie D: Sciences naturelles,
Fernando Fernández-Ramírez, and Rosa M Bermúdez-Cruz, and Cecilia Montañez
September 1965, Biochimica et biophysica acta,
Fernando Fernández-Ramírez, and Rosa M Bermúdez-Cruz, and Cecilia Montañez
March 1967, Biochimica et biophysica acta,
Fernando Fernández-Ramírez, and Rosa M Bermúdez-Cruz, and Cecilia Montañez
January 2004, Nucleic acids research,
Fernando Fernández-Ramírez, and Rosa M Bermúdez-Cruz, and Cecilia Montañez
March 1969, Journal of bacteriology,
Copied contents to your clipboard!