Overlapping regions of Caf20 mediate its interactions with the mRNA-5'cap-binding protein eIF4E and with ribosomes.
Overlapping regions of Caf20 mediate its interactions with the mRNA-5'cap-binding protein eIF4E and with ribosomes.
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DOI：
10.1038/s41598-021-92931-4
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发表时间：
2021-06-29
影响因子：
4.6
通讯作者：
Pavitt GD
中科院分区：
文献类型：
--
作者：
Nwokoye EC;AlNaseem E;Crawford RA;Castelli LM;Jennings MD;Kershaw CJ;Pavitt GD
翻译起始因子eIF 4 E通过与mRNA的5′端帽相互作用，在真核细胞中选择用于蛋白质合成的mRNA中起关键作用。Caf 20是在真核生物中发现的称为4 E-BP的蛋白质家族的成员，其与eIF 4G竞争与eIF 4 E的相互作用。Caf 20独立地与核糖体相互作用。因此，Caf 20通过知之甚少的机制调节mRNA选择过程。在这里，我们对Caf 20进行了无偏诱变，以确定Caf 20的哪些区域对于与eIF 4 E和核糖体的相互作用是重要的。Caf 20与eIF 4 E的结合完全依赖于与其他4 E-BP共享的典型基序。然而，与核糖体的结合被整个蛋白质的突变所削弱，这表明与eIF 4 E相互作用区域部分重叠的延伸的结合界面。通过使用化学交联，我们确定了一个潜在的核糖体表面上的核糖体相互作用区域，跨越大小亚基，并接近一个已知的相互作用位点eIF 3。Caf 20与核糖体结合的功能尚不清楚。
By interacting with the mRNA 5′ cap, the translation initiation factor eIF4E plays a critical role in selecting mRNAs for protein synthesis in eukaryotic cells. Caf20 is a member of the family of proteins found across eukaryotes termed 4E-BPs, which compete with eIF4G for interaction with eIF4E. Caf20 independently interacts with ribosomes. Thus, Caf20 modulates the mRNA selection process via poorly understood mechanisms. Here we performed unbiased mutagenesis across Caf20 to characterise which regions of Caf20 are important for interaction with eIF4E and with ribosomes. Caf20 binding to eIF4E is entirely dependent on a canonical motif shared with other 4E-BPs. However, binding to ribosomes is weakened by mutations throughout the protein, suggesting an extended binding interface that partially overlaps with the eIF4E-interaction region. By using chemical crosslinking, we identify a potential ribosome interaction region on the ribosome surface that spans both small and large subunits and is close to a known interaction site of eIF3. The function of ribosome binding by Caf20 remains unclear.
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DOI：
10.1073/pnas.2022756118
发表时间：
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