増殖細胞核抗原
増殖細胞核抗原(ぞうしょくさいぼうかくこうげん、英: proliferating cell nuclear antigen、略称: PCNA)は、真核生物細胞においてDNAポリメラーゼδのプロセシビティ因子として作用するDNAクランプであり、DNA複製に必要不可欠である。PCNAはホモ三量体を形成し、DNAを取り囲むことでプロセシビティを高め、DNA複製、DNA修復、クロマチンリモデリング、エピジェネティクスに関与するタンパク質をリクルートするための足場として機能する[5]。
多くのタンパク質は、PIP(PCNA-interacting peptide)ボックス[6]とAPIM(AlkB homologue 2 PCNA interacting motif)[7]という2つのPCNA相互作用モチーフを介してPCNAと相互作用する。PIPボックスを介してPCNAに結合するタンパク質が主にDNA複製に関与しているのに対し、APIMを介してPCNAに結合するタンパク質は主に遺伝毒性ストレスとの関係で重要である[8]。
機能
[編集]PCNA遺伝子にコードされるPCNAタンパク質は核内に存在し、DNAポリメラーゼδ(Pol δ)のコファクターである。PCNAはホモ三量体の形で機能し、DNA複製時にリーディング鎖合成のプロセシビティの向上を補助する。また、DNA損傷に応答してPCNAはユビキチン化され、RAD6依存的DNA修復経路に関与する。この遺伝子には同一のタンパク質をコードする2種類の転写産物が見つかっている。この遺伝子の偽遺伝子が4番染色体とX染色体に存在する[9]。
DNA合成時における核内での発現
[編集]PCNAはもともと、細胞周期のDNA合成期(S期)に細胞核で発現している抗原として同定された[10]。タンパク質の配列の一部が解析され、それをもとにcDNAクローンが単離された[11]。PCNAはPol δのDNAへの保持を補助する。PCNAは複製因子C(RFC)の作用によってDNAへのクランプを形成する[12][13]。RFCはAAA+ファミリーのATPアーゼのヘテロ五量体型のメンバーである。PCNAの発現は、転写因子E2Fを含む複合体の制御下にある[14][15]。
DNA修復における役割
[編集]DNAポリメラーゼδやεはDNA修復時に除去された損傷DNA鎖の再合成に関与しているため、PCNAはDNA合成とDNA修復の双方に重要である[16][17]。
PCNAは複製後修復(PRR)と呼ばれるDNA損傷トレランス経路にも関与している[18]。PRRには2つのサブ経路が存在し、損傷乗り越え(translesion)経路は損傷したDNA塩基を活性部位に取り込むことができる特殊なDNAポリメラーゼによって行われ(通常の複製ポリメラーゼは停止してしまう)、鋳型乗り換え(template switch)経路では相同組換え装置のリクルートによって損傷部位の迂回が行われると考えられている。PCNAはこれらの経路の活性化や、どちらの経路が利用されるかの選択に重要である。PCNAはユビキチン化による翻訳後修飾が行われる[19]。PCNAのリジン164番のモノユビキチン化は損傷乗り越え経路を活性化する。このモノユビキチンに対して非典型的なリジン63番連結型のポリユビキチン化が行われると、鋳型乗り換え経路が活性化されると考えられている[19]。さらに、PCNAのリジン164番(と程度は低いもののリジン127番)のSUMO化は鋳型乗り換え経路を阻害する[19]。SUMO化されたPCNAはSrs2と呼ばれるDNAヘリカーゼをリクルートし[20]、相同組換えの開始に重要なRad51ヌクレオタンパク質フィラメントを破壊することでこの拮抗的な作用を示す。
相互作用
[編集]PCNAには、DNAポリメラーゼ、クランプローダー、フラップエンドヌクレアーゼ、DNAリガーゼ、トポイソメラーゼ、ライセンス化因子、E3ユビキチンリガーゼ、E2 SUMO結合酵素、ヘリカーゼ(ATPアーゼ)、ミスマッチ修復酵素、塩基除去修復酵素、ヌクレオチド除去修復酵素、ポリ(ADP-リボース)ポリメラーゼ、ヒストンシャペロン、クロマチンリモデリング因子、ヒストンアセチル化酵素、ヒストン脱アセチル化酵素、DNAメチルトランスフェラーゼ、姉妹染色分体接着因子、プロテインキナーゼ、細胞周期調節因子、アポトーシス関連因子など、多くのタンパク質が結合する[21]。
より具体的には、次の挙げる因子との相互作用が示されている。
- ALKBH2[7]
- ANXA2[22]
- CAF-1[23][24][25]
- CDC25C[26]
- CHTF18[22]
- CCND1[27][28]
- CCNO[22][29]
- CDK4[28][30]
- CDKN1C[31]
- DNMT1[32][33][34]
- EP300[35]
- ESCO2[36]
- FBH1[37]
- FEN1[38][39][40][41][42][43][44]
- GADD45A[45][46][47][48][49]
- GADD45G[50][51]
- GTF2I[7]
- HDAC1[52]
- HUS1[53]
- ING1[54]
- KCTD13[55]
- Ku70[22][56]
- Ku80[22][56][57]
- MCL1[58]
- MSH3[22][59][60]
- MSH6[22][59][60]
- MUTYH[61]
- P21[31][40][44][62][63][64][65][66]
- PCLAF[44]
- POLD2[67]
- POLD3[22][68]
- POLDIP2[69]
- POLH[70]
- POLL[71][72][73]
- RFC1[22][62][74][75][76]
- RFC2[22][77][78]
- RFC3[22][79]
- RFC4[22][77]
- RFC5[22][75][77]
- UBC[80][81][82]
- WRN[83][84]
- XPA[85]
- XRCC1[86]
- YBX1[87]
- ZRANB3[88]
利用
[編集]PCNAに対する抗体またはKi-67と呼ばれるモノクローナル抗体が、星細胞腫などさまざまな新生物のグレーディングに利用される。これらは診断や予後の判定に利用できる。抗体標識によるPCNAの核内分布のイメージングは、細胞周期のS期の初期、中期、後期を区別するために用いることができる[89]。しかしながら、抗体を用いる際の重要な制限として、細胞を固定する必要があるためにアーティファクトが生じる可能性がある。
一方、生細胞における複製と修復のダイナミクスの研究は、PCNAと蛍光タンパク質などとの融合タンパク質の導入によって行われる場合がある。また、トランスフェクションの必要性やトランスフェクションが困難であったり細胞の寿命が短いといった問題を回避するために、細胞透過性ペプチドによる複製・修復マーカーが利用される場合がある。これらのペプチドは、生きた組織においてin situで使用でき、複製中の細胞と修復中の細胞を区別することもできるという利点がある[90]。
PCNAは、がん治療における治療標的としての可能性がある[91]。
出典
[編集]- ^ a b c GRCh38: Ensembl release 89: ENSG00000132646 - Ensembl, May 2017
- ^ a b c GRCm38: Ensembl release 89: ENSMUSG00000027342 - Ensembl, May 2017
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関連文献
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関連項目
[編集]外部リンク
[編集]- PCNA - MeSH・アメリカ国立医学図書館・生命科学用語シソーラス
- “ANA: Cell cycle related (Mitotic): PCNA type 1 and type 2 Antibody Patterns”. Antibody Patterns.com. 2008年4月15日閲覧。
- Dan Krotz. “Structure of a clamp–loader complex”. Advanced Light Source News. Lawrence Berkeley National Laboratory. 2008年4月15日閲覧。
- Overview of all the structural information available in the PDB for UniProt: P12004 (Proliferating cell nuclear antigen) at the PDBe-KB.