The C nucleotide at the mature 5' end of the Escherichia coli proline tRNAs is required for the RNase E cleavage specificity at the 3' terminus as well as functionality.
The C nucleotide at the mature 5' end of the Escherichia coli proline tRNAs is required for the RNase E cleavage specificity at the 3' terminus as well as functionality.
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DOI：
10.1093/nar/gkab1260
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发表时间：
2022-02-22
影响因子：
14.9
通讯作者：
Kushner SR
中科院分区：
文献类型：
--
作者：
Mohanty BK;Maples V;Kushner SR
在大肠杆菌中，脯氨酸tRNA的3′-成熟是在CCA决定簇后立即通过一步RNase E内切核酸裂解发生的。这种加工途径与其他tRNA的3′-末端成熟不同，避免了广泛使用的3′ → 5′核酸外切加工、3′-聚腺苷酸化和随后的降解。在这里，我们表明，在成熟的proK和proL tRNA的5′-末端的胞嘧啶（C）是必需的RNA酶E切割后立即CCA决定簇和它们的功能。因此，将proL和proK tRNA成熟5′-末端的C核苷酸改变为更常见的G核苷酸导致RNA酶E切割CCA决定簇下游1-4个核苷酸。此外，5′-修饰的突变体tRNA需要RNase T和RNase PH才能进行3′-成熟，并成为多聚腺苷酸化和降解的底物。引人注目的是，由于脯氨酰-tRNA-合成酶的识别元件的变化，5′-修饰的脯氨酸tRNA的氨酰化被阻断。pheV 5′-成熟末端从G到C核苷酸的类似修饰不支持细胞活力。这一结果为成熟tRNA的第一个核苷酸在其加工和功能中的重要性提供了额外的支持。
Proline tRNA 3′-maturation in Escherichia coli occurs through a one-step RNase E endonucleolytic cleavage immediately after the CCA determinant. This processing pathway is distinct from the 3′-end maturation of the other tRNAs by avoiding the widespread use of 3′ → 5′ exonucleolytic processing, 3′-polyadenylation and subsequent degradation. Here, we show that the cytosine (C) at the mature 5′-terminus of the proK and proL tRNAs is required for both the RNase E cleavage immediately after the CCA determinant and their functionality. Thus, changing the C nucleotide at the mature 5′-terminus of the proL and proK tRNAs to the more common G nucleotide led to RNase E cleavages 1–4 nucleotides downstream of the CCA determinant. Furthermore, the 5′-modified mutant tRNAs required RNase T and RNase PH for their 3′-maturation and became substrates for polyadenylation and degradation. Strikingly, the aminoacylation of the 5′-modified proline tRNAs was blocked due to the change in the recognition element for prolyl-tRNA-synthetase. An analogous modification of the pheV 5′-mature terminus from G to C nucleotide did not support cell viability. This result provides additional support for the importance of first nucleotide of the mature tRNAs in their processing and functionality.
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影响因子：
14.9
作者：
Mohanty, Bijoy K.;Agrawal, Ankit;Kushner, Sidney R.
通讯作者：
Kushner, Sidney R.
编码tRNA（LEU5）的大肠杆菌Leux tRNA转录物的处理需要3' - > 5'驱核酸酶核酸酶多核苷酸磷酸化酶或RNase P，以去除独立的转录终结剂。
影响因子：
14.9
作者：
Mohanty BK;Kushner SR
通讯作者：
Kushner SR
DOI：
10.1073/pnas.120163297
发表时间：
2000-06-06
影响因子：
11.1
作者：
Datsenko, KA;Wanner, BL
通讯作者：
Wanner, BL
影响因子：
5.6
作者：
Dittmar, KA;Mobley, EM;Pan, T
通讯作者：
Pan, T
DOI：
10.1073/pnas.92.15.6883
发表时间：
1995-07-18
影响因子：
11.1
作者：
LI, ZW;DEUTSCHER, MP
通讯作者：
DEUTSCHER, MP