9.10
真核細胞では、RNA ポリメラーゼによって作られた転写物が核から出る前に修飾および処理されます。 プロセシングされていない RNA は、成熟 mRNA と区別するために、前駆体 mRNA またはプレ mRNA と呼ばれます。
約 20 ~ 40 個のリボヌクレオチドが RNA ポリメラーゼによって結…
真核細胞では、新たに転写されたmRNAはprecursor mRNA(pre-mRNA)と呼ばれます。各プレmRNAは、2つの重要な修飾を受けています。1つは分子の5プライム末端(キャップ)にあり、もう1つは分子の3プライム末端(テール)にあります。
ファイブプライムキャップは、修飾されたグアニンヌクレオチドである単一の7-メチルグアノシンで構成されており、このヌクレオチドは三リン酸結合によってpre-mRNAの最初のヌクレオチドに結合しています。
プレmRNA転写産物の3プライムエンドに向かうヌクレオチドの特定のセット(通常はA、A、U、A、A、A)は、ポリアデニル化シグナルと呼ばれます。それは3プライム末端処理機構を動員し、エンドリボヌクレアーゼに3プライム切断部位で転写産物を切断するように指示します。
次に、別の酵素であるポリアデニル酸ポリメラーゼ(PAP)が、転写産物の3プライム末端に200個もの長いアデニンヌクレオチドを追加します。
5プライムキャップと3プライムポリAテールは、転写産物の末端をエキソヌクレアーゼによる分解から保護します。また、スリープライムテールは、mRNA転写産物が核を離れる準備ができていることを輸送分子に信号を送ります。核の外側では、5プライムキャップがリボソームが転写産物に付着し、翻訳を開始するのを助けます。
View the full transcript and gain access to JoVE Core videos
Q1: What is the 5' cap and why is it added to pre-mRNA?
The 5' cap is a 7-methylguanosine, a modified guanine nucleotide attached to the first nucleotide of pre-mRNA via a triphosphate linkage. It protects the transcript from degradation by exonucleases and helps the ribosome attach during translation. The cap also helps cells distinguish mRNA from other RNA types.
Q2: How does the poly-A tail protect mRNA and signal its readiness for translation?
Polyadenylate polymerase adds approximately 200 adenine nucleotides to the 3' end of pre-mRNA, forming the poly-A tail. This tail protects mRNA from degradation by exonucleases in the cytoplasm and signals to transport molecules that the transcript is ready to exit the nucleus for translation.
Q3: What is a polyadenylation signal and how does it direct 3' end processing?
A polyadenylation signal is a specific sequence of nucleotides, typically A, A, U, A, A, A, located toward the 3' end of pre-mRNA. It recruits the 3' end processing machinery and directs an endoribonuclease to cut the transcript at the 3' cleavage site, allowing polyadenylate polymerase to add the poly-A tail.
Q4: What is the difference between exons and introns in pre-mRNA?
Exons are RNA sequences that remain in the mature transcript and are expressed as proteins. Introns are portions removed during splicing by the spliceosome. Interestingly, a single RNA segment can function as an exon in one cell type and an intron in another, enabling alternative rna splicing to produce multiple proteins from one gene.
Q5: When does the 5' cap get added during transcription?
The 5' cap is added early during transcription, once approximately 20-40 ribonucleotides have been joined together by RNA polymerase. A group of enzymes replaces the 5' phosphate with modified 7-methyl guanosine, which helps the cell recognize mRNA and prepares it for subsequent translation.
Q6: How do the 5' cap and 3' poly-A tail work together to protect mRNA?
Both the 5' cap and 3' poly-A tail protect mRNA ends from degradation by exonucleases. The 5' cap marks the transcript as mRNA and facilitates ribosome binding, while the 3' tail signals nuclear export and protects the transcript in the cytoplasm, ensuring the mRNA remains stable for translation.
Q7: What happens to pre-mRNA after the 5' cap and poly-A tail are added?
After both modifications are complete, the mature mRNA exits the nucleus for translation. The 5' cap and 3' poly-A tail signal to transport molecules that the transcript is ready to leave the nucleus. Outside the nucleus, the 5' cap helps the ribosome attach to initiate protein synthesis.