李園園，郭磊，韓之明,3

綜 述

NEK家族在細(xì)胞周期調(diào)控中的作用

李園園1，郭磊2，韓之明1,3

1. 中國科學(xué)院動(dòng)物研究所，干細(xì)胞與生殖生物學(xué)國家重點(diǎn)實(shí)驗(yàn)室，北京 100101 2. 廣東省第二人民醫(yī)院生殖醫(yī)學(xué)中心，生殖力保護(hù)實(shí)驗(yàn)室，廣州 510317 3. 北京干細(xì)胞與再生醫(yī)學(xué)研究院，北京 100101

NIMA相關(guān)激酶(NIMA-related kinases, NEKs)是絲氨酸/蘇氨酸激酶，在細(xì)胞周期調(diào)控中發(fā)揮著重要的作用，參與了中心體分離、紡錘體組裝、染色質(zhì)凝集、核膜破裂、紡錘體組裝檢驗(yàn)點(diǎn)信號(hào)、胞質(zhì)分裂、纖毛形成及DNA損傷反應(yīng)等多種細(xì)胞活動(dòng)。本文結(jié)合近年來在該激酶家族的相關(guān)研究，從NEK家族的組成、結(jié)構(gòu)特征及其在有絲分裂和減數(shù)分裂過程中的作用等多個(gè)方面展開綜述，以期為進(jìn)一步研究NEKs在細(xì)胞周期調(diào)控中的作用提供重要基礎(chǔ)，也為腫瘤的臨床診斷和治療提供理論依據(jù)。

NIMA相關(guān)激酶；有絲分裂；減數(shù)分裂

細(xì)胞是生命活動(dòng)的基本單位。細(xì)胞周期是一個(gè)非常復(fù)雜和精細(xì)的調(diào)節(jié)過程，該過程受到細(xì)胞內(nèi)外各種因素的精密調(diào)控，細(xì)胞周期的紊亂與許多疾病的發(fā)生發(fā)展相關(guān)。研究顯示，許多蛋白激酶家族，如細(xì)胞周期蛋白依賴性激酶(cyclin-dependent kinases, CDK)、Aurora激酶、Polo樣激酶(polo-like kinase, PLK)和NIMA相關(guān)激酶(NIMA-related kinases, NEKs)，都參與了細(xì)胞周期調(diào)控的過程。近年的研究發(fā)現(xiàn)，NEK家族蛋白在細(xì)胞周期調(diào)控的過程中扮演了重要的角色，參與中心體復(fù)制和分離、紡錘體形成、染色體在赤道板上的排列、紡錘體檢驗(yàn)點(diǎn)(spindle assembly checkpoint，SAC)調(diào)控、纖毛形成及DNA損傷反應(yīng)(DNA damage response, DDR)等多種細(xì)胞活動(dòng)。本文主要綜述了NEK家族成員的生物學(xué)特性及其在細(xì)胞周期調(diào)控中的作用，同時(shí)對(duì)NEK家族的未來研究方向進(jìn)行了探討，以期讓相關(guān)科研人員更充分、更全面地了解NEK家族的研究進(jìn)展，為進(jìn)一步研究其在細(xì)胞周期調(diào)控中的作用提供有力的支撐，也為深入了解腫瘤發(fā)生機(jī)制及抗腫瘤藥物設(shè)計(jì)提供研究基礎(chǔ)。

1 NEK家族及其生物學(xué)特性

1.1 NEK家族的發(fā)現(xiàn)

NIMA (never in mitosis A)最早是在對(duì)曲霉屬真菌的有絲分裂突變體的研究中發(fā)現(xiàn)的[1,2]。20世紀(jì)80年代中期，Osmani等[3]通過調(diào)控基因的mRNA表達(dá)水平證明參與了曲霉有絲分裂的G2/M期轉(zhuǎn)換。進(jìn)一步的研究證明，的過表達(dá)可以促進(jìn)有絲分裂的提前發(fā)生，有絲分裂過程中NIMA與CDK1-cyclin B復(fù)合體是同等重要的調(diào)節(jié)因子[4,5]。在對(duì)曲霉的研究中發(fā)現(xiàn)，NIMA的降解是細(xì)胞完成正確的有絲分裂進(jìn)程所必需的[6]。一系列的研究表明，NIMA激酶在曲霉和酵母()中參與了染色質(zhì)凝集、紡錘體組裝和胞質(zhì)分裂等多個(gè)細(xì)胞周期過程[7～11]。20世紀(jì)90年代初期，Letwin等[12]從小鼠()中分離出，發(fā)現(xiàn)編碼一種與NIMA相關(guān)的蛋白激酶，在結(jié)構(gòu)、組成和表達(dá)上與NIMA存在較高的一致性，從而提出了在哺乳動(dòng)物中可能存在一個(gè)基因家族。隨后的研究發(fā)現(xiàn)了小鼠和人()的細(xì)胞中均存在與相關(guān)的基因，證明了高等哺乳動(dòng)物確實(shí)存在NEK家族[13,14]。研究已證明，NEKs存在于多種生物體中，從原生生物如衣藻()[15]、瘧原蟲()[16]等到多細(xì)胞真核生物如果蠅()[17]、非洲爪蟾()[18]、小鼠[13]和人[14]。

1.2 NEK家族成員的結(jié)構(gòu)特征

人類NEK家族由11種NIMA相關(guān)激酶組成[19,20]，這些激酶具有與曲霉NIMA相似的氨基末端催化區(qū)域，是含有典型的絲氨酸/蘇氨酸激酶序列的高度保守的激酶結(jié)構(gòu)域，其氨基末端和羧基末端的調(diào)節(jié)結(jié)構(gòu)域在序列組成和長度上有顯著差異。一般來講，NEK家族的氨基末端激酶區(qū)域是中度保守的，與NIMA的激酶區(qū)域的氨基酸序列有40%～50%的同源性。NEK10的激酶區(qū)域位于整個(gè)氨基酸序列的中段，與NEK家族典型的氨基末端催化區(qū)域不同。在NEK家族中，人NEK2和NIMA的同源性最高，能達(dá)到44%[21]。除此之外，NEK6和NEK7的激酶區(qū)域的序列一致性達(dá)到了85%以上[22]。人類NEK家族的催化區(qū)域均含有一個(gè)His-Arg-Asp(HRD)基序，在激活環(huán)中都有一個(gè)絲氨酸/蘇氨酸殘基，而這個(gè)殘基很可能是激活修飾的作用位點(diǎn)。在一些NEK家族成員中，這個(gè)殘基是自磷酸化的，而其他成員則是通過一個(gè)上游激酶進(jìn)行磷酸化修飾的[23～26]。就磷酸化識(shí)別序列而言，NIMA的第3位殘基具有對(duì)苯丙氨酸的強(qiáng)烈偏好[27]，人類NEK家族也具有相似的偏好，例如NEK2和NEK6的第3位殘基更喜歡疏水殘基，尤其偏愛苯丙氨酸或亮氨酸[28,29]。

NEK家族成員具有保守的氨基末端催化區(qū)域，而羧基末端區(qū)域在長度、序列和結(jié)構(gòu)上都存在很大差異(圖1)。其常見的特點(diǎn)就是寡聚化序列，通常是一種卷曲螺旋結(jié)構(gòu)，可通過自磷酸化而被激活。一般而言，自磷酸化通常是在激酶結(jié)構(gòu)域的激活環(huán)內(nèi)進(jìn)行，但是也可發(fā)生在蛋白質(zhì)的其他區(qū)域，例如NEK8和NEK9羧基末端的非催化區(qū)域可以通過自磷酸化調(diào)控自身的定位和激活[23,30]。研究發(fā)現(xiàn)，包括曲霉NIMA和脊椎動(dòng)物NEK2在內(nèi)的幾種NEKs均顯示在非催化區(qū)域內(nèi)存在靶向蛋白質(zhì)降解的破壞基序[6,31]，例如NEK2含有一個(gè)KEN (Lys-Glu-Asn)- box和羧基末端MR (methionine-arginine dipeptide)- tail，均能被后期促進(jìn)復(fù)合物/環(huán)狀體(anaphase- promoting complex/cyclosome, APC/C)所識(shí)別，其中MR-tail還可介導(dǎo)NEK2與APC/C的核心亞基CDC20直接作用，從而導(dǎo)致NEK2以一種不依賴于紡錘體組裝檢驗(yàn)點(diǎn)的方式進(jìn)行降解[32]。在NEK家族中，NEK6和NEK7僅由一個(gè)催化區(qū)域和短的氨基末端延伸區(qū)域組成[33,34]，而后者可能與底物識(shí)別有關(guān)[35]。NEK6和NEK7是NEK9的下游激酶，可以和NEK9蛋白中RCC1域和coiled-coil域之間的一個(gè)序列結(jié)合[25]。

圖1 人NEK家族的結(jié)構(gòu)特征

2 NEK家族在細(xì)胞周期調(diào)控中的作用

作為蛋白激酶，NEK家族參與了細(xì)胞周期、細(xì)胞分裂、纖毛形成和DNA損傷反應(yīng)等多種細(xì)胞活動(dòng)(表1)。人和哺乳動(dòng)物NEK家族在細(xì)胞有絲分裂和減數(shù)分裂過程中的作用主要有以下幾個(gè)方面。

2.1 NEK家族在有絲分裂中的作用

的過表達(dá)可以誘導(dǎo)處于細(xì)胞周期任何階段的曲霉細(xì)胞、酵母細(xì)胞、非洲爪蟾卵母細(xì)胞或人類細(xì)胞進(jìn)入有絲分裂[82,83]，研究發(fā)現(xiàn)，人類NEK家族參與細(xì)胞周期進(jìn)程和分化過程中的多個(gè)事件。在有絲分裂中，NEK2、NEK6、NEK7和NEK9相互配合調(diào)控雙極紡錘體的形成、染色質(zhì)凝集、核膜破裂和胞質(zhì)分裂等。NEK3除參與調(diào)控有絲分裂外，還可促進(jìn)催乳素依賴性信號(hào)傳導(dǎo)[45]，而NEK1、NEK4、NEK5、NEK7、NEK8、NEK10和NEK11均與DNA損傷應(yīng)答有關(guān)。

2.1.1 有絲分裂起始

有絲分裂的起始和退出是由CDK1、cyclins、有絲分裂相關(guān)激酶和磷酸酶驅(qū)動(dòng)的細(xì)胞周期轉(zhuǎn)換。在高等真核生物中，有絲分裂的起始導(dǎo)致多個(gè)細(xì)胞結(jié)構(gòu)的改變，例如中心體分離、微管生長和收縮、核膜破裂以及染色質(zhì)凝集等[84]。盡管沒有研究證明NEK家族是有絲分裂起始所必需的，但是已確定NEK2、NEK6、NEK7和NEK9參與調(diào)控了細(xì)胞從間期進(jìn)入M期的中心體的分離、紡錘體的組裝、核孔復(fù)合物的去組裝和核膜破裂等。

表1 人和哺乳動(dòng)物NEK家族的亞細(xì)胞定位和功能

研究發(fā)現(xiàn)，一些NEK家族成員在從真菌到人類的微管組織中心均有定位[9,17,85～87]。在人類細(xì)胞中，NEK2作為中心體的核心組分，參與調(diào)控中心體的分離[41,88,89]。在有絲分裂間期，兩個(gè)中心粒由一些蛋白質(zhì)連接體結(jié)合在一起，而該連接體是由卷曲螺旋蛋白組成的，包括C-Nap1、rootletin、Cep68、centlein和LRRC45，而NEK2不僅可通過磷酸化連接蛋白[90～94]和中心粒相關(guān)蛋白GAS2L1[95,96]，還可通過失活驅(qū)動(dòng)蛋白KIFC3[97]，共同調(diào)控有絲分裂前期的中心體分離和雙極紡錘體形成。在有絲分裂間期，NEK2與蛋白激酶MST-2和磷酸酶PP1形成三聚體結(jié)構(gòu)，維持在一個(gè)去磷酸化的失活狀態(tài)。當(dāng)有絲分裂啟動(dòng)時(shí)，PLK1可通過磷酸化MST-2破壞這種結(jié)構(gòu)，導(dǎo)致NEK2的激活。除此之外，NEK2也可通過自磷酸化而被激活[98]。在有絲分裂過程中，NEK5與NEK2的定位模式相似。人基因的敲降導(dǎo)致分裂間期NEK2減少、中心粒周圍物質(zhì)(pericentriolar material, PCM)缺失、微管生長緩慢以及中心體連接蛋白rootletin被過度募集到中心體上，從而導(dǎo)致中心體的過早分離，分離的中心體之間相對(duì)較接近[50]，這個(gè)現(xiàn)象與過表達(dá)人基因的結(jié)果是一致的[41,91]，而且同時(shí)敲降和基因后中心體的過早分離被加重。我們推測(cè)，NEK5可能與NEK2協(xié)同調(diào)控中心體的分離。

研究發(fā)現(xiàn)，在有絲分裂的G1期和S期，NEK7可通過調(diào)控PCM的募集促進(jìn)中心體的復(fù)制[99]。人基因的敲降導(dǎo)致PCM組分和原中心粒組裝相關(guān)蛋白PLK4、CPAP、SAS-6以及STIL不能被募集到中心體，從而調(diào)控中心體的復(fù)制[100]，而人基因和基因的過表達(dá)能夠誘導(dǎo)額外的中心體形成[101]。在有絲分裂中，人、和基因的敲降導(dǎo)致前期中心體的分離失敗、分裂中期形成脆弱的紡錘體、紡錘體兩極的距離減小以及微管密度降低[23,56,66]。事實(shí)上，對(duì)于這些紡錘體的缺陷最簡單的解釋是中心體和紡錘體兩極的微管成核作用減少。研究顯示，NEK9能與啟動(dòng)微管成核的γ-tubulin環(huán)狀復(fù)合體(γ-tubulin ring complex, γ-TuRC)的多個(gè)組分互作，如磷酸化γ-TuRC的銜接蛋白NEDD1[73,102]，后者的激活促進(jìn)了γ-tubulin被募集到中心體上，而的缺失會(huì)導(dǎo)致紡錘體組裝延遲、雙極紡錘體的形成減少和微管結(jié)構(gòu)異常[102]。此外，NEK6和NEK7均定位到紡錘體兩極，NEK6在有絲分裂的中期和后期定位到紡錘體微管上[56]，NEK7可將γ-tubulin募集到紡錘體的兩極[66]。研究結(jié)果提示，這些激酶對(duì)微管成核的調(diào)控可能不僅是通過紡錘體兩極和紡錘體本身，還有可能是通過augmin復(fù)合體將γ-TuRCs募集到紡錘體的兩極[103]。除此之外，這些激酶調(diào)控紡錘體形成的另一種途徑可能是通過磷酸化微管相關(guān)蛋白進(jìn)行的，例如Eg5作為一種驅(qū)動(dòng)蛋白，參與了有絲分裂雙極紡錘體的形成和維持過程，而Eg5被募集到紡錘體微管上的過程依賴于CDK1對(duì)Eg5的磷酸化作用[104,105]。研究發(fā)現(xiàn)，NEK6也可磷酸化Eg5[106]，這一發(fā)現(xiàn)有助于闡明NEK6或NEK9在雙極紡錘體的形成和維持中的作用[23,106]。另一項(xiàng)研究顯示，EML4作為一種促進(jìn)微管穩(wěn)定性的微管相關(guān)蛋白參與微管動(dòng)力學(xué)的調(diào)控，NEK6和NEK7可通過磷酸化EML4降低其與微管的親和力，從而促進(jìn)染色體中板聚合[107]。NEK6和NEK7還可以直接將微管蛋白磷酸化，這一發(fā)現(xiàn)提示NEK6和NEK7可能通過磷酸化微管蛋白直接參與微管動(dòng)力學(xué)的調(diào)控[56]。這些研究均表明，NEK6、NEK7和NEK9在紡錘體的形成中發(fā)揮了重要作用。

NEK2、NEK6、NEK7和NEK9除影響紡錘體形成之外，也發(fā)揮其他的功能。例如，NEK2的剪接異構(gòu)體NEK2C定位在細(xì)胞核中，這可能與NEK2在細(xì)胞核中的功能有關(guān)[108]。研究顯示，Nup98是核孔復(fù)合體(nuclear pore complexes, NPCs)的組成成分，CDK1和NIMA可磷酸化Nup98，從而促進(jìn)Nup98從NPCs的解離。CDK1還可磷酸化NEK9的Ser869位點(diǎn)，進(jìn)而激活NEK9，而NEK6和NEK7可通過與激活的NEK9結(jié)合而被激活[23]。因此，我們推測(cè)NEKs也可能參與NPCs的解體和核膜破裂[109]。除此之外，NEK9還可與BICD2相互作用。而BICD2作為一種動(dòng)粒蛋白相關(guān)蛋白，在有絲分裂前期可與動(dòng)力蛋白結(jié)合，促進(jìn)核孔復(fù)合體的去組裝[110]。這些研究結(jié)果均表明，NEK家族在有絲分裂起始中發(fā)揮重要作用。

2.1.2 細(xì)胞周期檢驗(yàn)點(diǎn)

細(xì)胞周期阻滯可發(fā)生在細(xì)胞周期的G1/S、S期和G2/M期，是由內(nèi)源性因素(如停滯的復(fù)制叉)或者外源性因素(包括紫外線(UV)輻射、電離輻射(IR)、活性氧(ROS)和某些化療藥物)所造成的DNA損傷引起的。細(xì)胞周期由一系列的檢驗(yàn)點(diǎn)所監(jiān)控，當(dāng)DNA出現(xiàn)損傷時(shí)，這些檢驗(yàn)點(diǎn)蛋白被激活，進(jìn)而導(dǎo)致細(xì)胞周期的延遲或阻滯。檢驗(yàn)點(diǎn)的激活是由PIKK (phosphatidylinositol-3 kinase-related kinase)家族成員共濟(jì)失調(diào)毛細(xì)血管擴(kuò)張突變(ataxia telangiectasia mutated, ATM)蛋白和共濟(jì)失調(diào)毛細(xì)血管擴(kuò)張突變與相關(guān)(ataxia telangiectasia mutated andrelated, ATR)蛋白及其效應(yīng)激酶CHK1/2 (checkpoint kinase 1/2)啟動(dòng)的，ERK1/2 (extracellular signal- regulated kinase 1/2)和p38及其下游激酶MK2 (MAPK activated protein kinase 2)在細(xì)胞周期阻滯中也發(fā)揮重要作用。在NEK家族中，NEK2和NEK6作為DNA損傷反應(yīng)的靶點(diǎn)，是受DNA損傷抑制的[58,111]，而其他的NEK家族成員在DNA損傷修復(fù)中發(fā)揮重要作用。

在有絲分裂的G1/S和G2/M轉(zhuǎn)換中，NEK1在DNA損傷修復(fù)中起作用[112～115]。當(dāng)敲除的細(xì)胞暴露于IR和UV輻射時(shí)，CHK1和CHK2不能被激活。此外，NEK1的激活不依賴于ATM和ATR。這些研究結(jié)果提示，NEK1可能是作為損傷信號(hào)的獨(dú)立傳感器發(fā)揮作用。

研究發(fā)現(xiàn)，NEK2不僅可與SAC蛋白相互作用，還可促進(jìn)動(dòng)粒復(fù)合蛋白HEC1的Ser165位點(diǎn)磷酸化[116～118]。除此之外，在紊亂的染色體動(dòng)粒上可檢測(cè)到磷酸化HEC1 (Ser165)的表達(dá)，而HEC1可將MPS1和MAD1/MAD2復(fù)合體募集到動(dòng)粒上[119]。由此推測(cè)，NEK2可能參與紡錘體組裝檢驗(yàn)點(diǎn)SAC蛋白完整性的調(diào)控。

研究還發(fā)現(xiàn)，NEK8可通過RAD51蛋白和DNA損傷修復(fù)調(diào)控復(fù)制叉的穩(wěn)定性[71]，而NEK10和NEK11參與調(diào)控G2/M期的DNA損傷反應(yīng)檢驗(yàn)點(diǎn)。當(dāng)細(xì)胞暴露于UV輻射時(shí)，NEK10與MEK1、RAF1形成一個(gè)三聚體的結(jié)構(gòu)，NEK10可通過促進(jìn)MEK1的激活，進(jìn)而導(dǎo)致G2/M期阻滯和ERK1/2的磷酸化[75]，敲降人基因可以抑制MEK1和ERK1/2的磷酸化。當(dāng)發(fā)生DNA損傷和遺傳毒性應(yīng)激時(shí)，NEK11活性顯著增加，而當(dāng)抑制ATM和ATR激酶時(shí)，NEK11不能被激活[79,80]。當(dāng)細(xì)胞暴露于IR輻射時(shí)，ATR和ATM激活CHK1，CHK1的激活促進(jìn)NEK11和CDC25A的磷酸化，而NEK11的激活可進(jìn)一步磷酸化CDC25A，這一過程促進(jìn)SCF泛素連接酶復(fù)合物與CDC25A的結(jié)合，從而促進(jìn)CDC25A的降解，最終導(dǎo)致G2/M期阻滯[79]，使細(xì)胞有充足的時(shí)間進(jìn)行DNA修復(fù)，不會(huì)過早進(jìn)入有絲分裂。

2.1.3 胞質(zhì)分裂

胞質(zhì)分裂發(fā)生在細(xì)胞分裂后期姐妹染色單體分離之后, 是細(xì)胞周期和生物個(gè)體發(fā)育過程中的一個(gè)重要環(huán)節(jié), 直接關(guān)系到遺傳物質(zhì)和細(xì)胞質(zhì)組分能否在2個(gè)子細(xì)胞中正常分配。胞質(zhì)分裂是由許多亞細(xì)胞結(jié)構(gòu)和生物分子相互協(xié)調(diào)作用的結(jié)果。動(dòng)物細(xì)胞胞質(zhì)分裂過程主要包括分裂溝的定位、胞質(zhì)分裂結(jié)構(gòu)收縮的組裝、分裂溝的產(chǎn)生和收縮、分裂溝膜泡的融合以及中間體的形成和剪切。

在真核生物中，NEK家族也參與胞質(zhì)分裂的調(diào)控。在裂殖酵母中，Grallert等[11]發(fā)現(xiàn)FIN1在胞質(zhì)分裂中起重要作用。在果蠅中，NEK2定位在有絲分裂后期的中體上，它的過表達(dá)可導(dǎo)致actin和anillin在卵裂溝的形成部位發(fā)生錯(cuò)位[17]。人NEK2剪接異構(gòu)體NEK2B的敲降可導(dǎo)致細(xì)胞無法完成胞質(zhì)分裂而形成多核細(xì)胞[120]。NEK6和NEK7也定位在有絲分裂后期的中體上，在胞質(zhì)分裂中NEK6的激酶活性達(dá)到最大[56,66,106]。人或基因敲降的細(xì)胞可成功進(jìn)入中期，但不能完成胞質(zhì)分裂，而且人或的等位基因突變體細(xì)胞也經(jīng)常出現(xiàn)胞質(zhì)分裂的失敗[56,66]。研究還發(fā)現(xiàn)，來自小鼠敲除胚胎的胚胎成纖維細(xì)胞也表現(xiàn)出胞質(zhì)分裂失敗的缺陷[121]。除此之外，NEK6和NEK9還可介導(dǎo)與胞質(zhì)分裂有關(guān)的驅(qū)動(dòng)蛋白MKLP2和KIF14的定位和募集[122]。以上證據(jù)均表明，NEK家族可能通過胞質(zhì)分裂相關(guān)因子的定位和活性改變調(diào)控胞質(zhì)分裂[56,122]。

2.2 NEK家族在減數(shù)分裂中的作用

如上所述，NEK家族在有絲分裂過程中發(fā)揮重要的調(diào)節(jié)作用。減數(shù)分裂作為一種特殊的細(xì)胞分裂方式，是真核生物和二倍體生物有性生殖和配子產(chǎn)生所必需的。在減數(shù)分裂中，染色體的錯(cuò)誤分離有可能導(dǎo)致非整倍體受精卵或后代的產(chǎn)生。與有絲分裂相比，人們對(duì)NEK家族在減數(shù)分裂中的作用了解較少。近些年的研究發(fā)現(xiàn)，一些NEK家族成員，如NEK1、NEK2、NEK5、NEK9和NEK11，在減數(shù)分裂中也發(fā)揮重要的作用。

在哺乳動(dòng)物生殖細(xì)胞中，NEK1高表達(dá)，并參與減數(shù)分裂中紡錘體形成的調(diào)控[36]。在敲除小鼠的精母細(xì)胞和卵母細(xì)胞中，第一次減數(shù)分裂的紡錘體組裝和染色體排列異常，調(diào)控紡錘體動(dòng)力相關(guān)蛋白-肌球蛋白X (myosin X, MYO10)和α-adducin的定位和表達(dá)改變[64,123,124]。我們推測(cè)，NEK1可能通過與MYO10和α-adducin的相互作用調(diào)控紡錘體的形成。在小鼠卵母細(xì)胞中，NEK2是微管組織中心的組成成分，它的敲降導(dǎo)致第一次減數(shù)分裂紡錘體兩極的異常和染色體排列異常[42]，研究證明centrobin/Nip2是NEK2的作用底物，在微管組織中心發(fā)揮重要作用[125,126]，而且在卵母細(xì)胞中敲降與敲降的表型是一致的[42]。這些結(jié)果提示，NEK2可能通過磷酸化centrobin參與調(diào)控卵母細(xì)胞減數(shù)分裂I中紡錘體組裝。在小鼠精母細(xì)胞減數(shù)分裂過程中，NEK2可磷酸化染色質(zhì)結(jié)構(gòu)蛋白HMGA2，通過降低后者與DNA的親和力調(diào)控染色質(zhì)的凝集[127]。我們最近的一項(xiàng)研究發(fā)現(xiàn)，NEK5在減數(shù)分裂G2/M轉(zhuǎn)換過程中發(fā)揮了重要作用，在敲降的卵母細(xì)胞中MPF活性降低，導(dǎo)致了卵母細(xì)胞減數(shù)分裂恢復(fù)的失敗[51]。同時(shí)，我們還發(fā)現(xiàn)NEK5定位在MI～MII期紡錘體上，推測(cè)NEK5也可能參與減數(shù)分裂紡錘體的組裝。在敲降的小鼠卵母細(xì)胞中，紡錘體組裝和染色體排列異常，γ-tubulin在紡錘體兩極的定位異常，SAC被激活[128]。在小鼠卵母細(xì)胞中敲降影響了MI期紡錘體的遷移，導(dǎo)致卵母細(xì)胞的均等分裂[81]。上述研究結(jié)果表明，在生殖細(xì)胞中NEK1、NEK2、NEK5和NEK9等是保證減數(shù)分裂正常進(jìn)行和染色體正確分離的關(guān)鍵蛋白，其表達(dá)的改變會(huì)導(dǎo)致紡錘體組裝相關(guān)因子的定位和活性改變進(jìn)而干擾紡錘體組裝和減數(shù)分裂細(xì)胞周期進(jìn)程。

3 結(jié)語與展望

自發(fā)現(xiàn)以來，NEK家族一直是細(xì)胞生物學(xué)的研究熱點(diǎn)，研究證明NEK家族在細(xì)胞周期調(diào)控中發(fā)揮著關(guān)鍵的作用，但其在減數(shù)分裂中的功能和分子機(jī)制還有待于進(jìn)一步深入的研究。細(xì)胞周期高度有序的運(yùn)轉(zhuǎn)是通過G1/S期轉(zhuǎn)換、G2/M轉(zhuǎn)換和中/后期轉(zhuǎn)換等多個(gè)過程的調(diào)控來實(shí)現(xiàn)的。細(xì)胞周期紊亂是腫瘤發(fā)生的主要原因，細(xì)胞周期相關(guān)蛋白的表達(dá)異常在腫瘤細(xì)胞增殖中扮演著重要角色。因此，對(duì)NEK家族的生物學(xué)功能及其在細(xì)胞周期調(diào)控中作用的研究，不僅可以更深入地了解細(xì)胞周期過程及調(diào)控機(jī)制，還有助于闡明NEK家族在腫瘤發(fā)生發(fā)展中的作用機(jī)制，對(duì)腫瘤的臨床診斷和治療也具有重要意義。

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Roles of NEK family in cell cycle regulation

Yuanyuan Li1, Lei Guo2, Zhiming Han1,3

As a serine/threonine kinase, NIMA-related kinases (NEKs) play important roles in the regulation of cell cycle, and involve in several cellular activities such as centrosome separation, spindle assembly, chromatin condensation, nuclear envelope breakdown, spindle assembly checkpoint signaling, cytokinesis, cilia formation and DNA damage response. In this review, we summarize the component, structural characteristics and functions of NEK family in mitosis and meiosis based on the relevant researches in recent years, providing a reference for the further study on the roles of NEKs in the regulation of cell cycle and a theoretical basis for the clinical diagnosis and treatment of tumors.

NIMA-related kinases; mitosis; meiosis

2021-03-27;

2021-05-12

國家重點(diǎn)研發(fā)計(jì)劃資助項(xiàng)目(編號(hào)：2018YFC1004000，2019YFA0109900)和國家自然科學(xué)基金項(xiàng)目(編號(hào)：31970509)資助[Supported by the National Key R&D Program of China (Nos. 2018YFC1004000, 2019YFA0109900), and the National Natural Science Foundation of China (No. 31970509)]

李園園，博士，專業(yè)方向：發(fā)育生物學(xué)。E-mail: liyuanyuan891116@163.com

韓之明，博士，副研究員，專業(yè)方向：發(fā)育生物學(xué)。E-mail: hanzm@ioz.ac.cn

10.16288/j.yczz.20-421

2021/6/25 13:16:28

URI: https://kns.cnki.net/kcms/detail/11.1913.R.20210625.1123.002.html

(責(zé)任編委: 史慶華)