Le Derout Jacques, Folichon Marc, Briani Federica, Dehò Gianni, Régnier Philippe, Hajnsdorf Eliane
UPR 9073 du CNRS, Institut de Biologie Physico-Chimique, 13 Rue Pierre et Marie Curie, 75005 Paris, France.
Nucleic Acids Res. 2003 Jul 15;31(14):4017-23. doi: 10.1093/nar/gkg456.
Polyadenylation plays an important role in RNA degradation in bacterial cells. In Escherichia coli, exoribonucleases, mostly RNase II and polynucleotide phosphorylase, antagonize the synthesis of poly(A) tails by poly(A) polymerase I (PAP I). In accordance with earlier observations showing that only a small fraction of bacterial RNA is polyadenylated, we demonstrate here that approximately 10% of rpsO mRNA harbors short oligo(A) tails ranging from one to five A residues in wild-type cells. We also compared the length, frequency and distribution of poly(A) tails at the 3'-end of rpsO transcripts in vivo in the presence and absence of Hfq, a host factor that in vitro stimulates the activity of PAP I, and found that Hfq affects all three parameters. In the hfq(+) strain the average length of oligo(A) tails and frequency of polyadenylated transcripts was higher than in the hfq(-) strain and a smaller proportion of tails was found at the 3' end of transcripts terminated at the Rho- independent terminator. Our data led us to the conclusion that Hfq is involved in the recognition of 3' RNA extremities by PAP I.
聚腺苷酸化在细菌细胞的RNA降解中起着重要作用。在大肠杆菌中,外切核糖核酸酶,主要是核糖核酸酶II和多核苷酸磷酸化酶,会拮抗聚腺苷酸聚合酶I(PAP I)合成聚(A)尾巴。与早期观察结果一致,即只有一小部分细菌RNA被聚腺苷酸化,我们在此证明,在野生型细胞中,约10%的rpsO mRNA带有长度从1到5个A残基不等的短寡聚(A)尾巴。我们还比较了在有和没有Hfq(一种在体外刺激PAP I活性的宿主因子)存在的情况下,体内rpsO转录本3'端聚(A)尾巴的长度、频率和分布,发现Hfq会影响所有这三个参数。在hfq(+)菌株中,寡聚(A)尾巴的平均长度和聚腺苷酸化转录本的频率高于hfq(-)菌株,并且在由不依赖Rho的终止子终止的转录本3'端发现的尾巴比例较小。我们的数据使我们得出结论,Hfq参与了PAP I对3' RNA末端的识别。