细胞周期及其控制

细胞周期(cell cycle)是指细胞从前一次分裂结束到下一次分裂结束这一间隔时期而言。在这个过程中包括两个阶段——细胞分裂间期和细胞分裂期。在细胞周期中,细胞要经历一系列的有规律的变化——形态学的和生理生化的。这些变化与生物体的生长发育和新陈代谢有着极为密切的关系。所以细胞周期的研究,不仅具有重要的理论意义,而且在生产实践中非常重要。本文着重讨论一下有关细胞周期的划分、细胞周期各个时期的生化特点和控制的近代观念。     

真核生物细胞周期的调控

近年来细胞周期调控已成为分子生物学研究的热点之一。本文对酵母、动物和植物的细胞分裂及细胞周期调控的研究进展作一简要综述，并对酵母和动物的细胞周期调控与植物的相应机制进行比较．     

磷酸饥饿诱导大肠杆菌同步化

用改良的Ｋｅｐｅｓ磷酸饥饿法诱导大肠杆菌Ｋ—１２ＡＭ１２６４同步化生长，细胞经８轮同步化步骤后可在磷酸不限制培养基中自由生长保持２～３次同步化细胞周期。Ｋ—１２ＡＭ１２６４大肠杆菌的细胞增倍和分化周期分别为５５和１５ｍｉｎ，同步化细胞周期可分达３个时相。细胞分裂期（Ｐ），细胞分裂和染色体复制起始间期（Ｑ）以及染色体复制起始和细胞分裂间期（Ｒ）。Ｒ期又可分Ｒ１和Ｒ２两个亚期。在Ｒ１亚期胸腺呼啶掺入ＤＮＡ的速度增加，在Ｒ２亚期掺入速度保持恒定。Ｒ１和Ｒ２期分别为１５和１０ｍｉｎ。     

洋葱根尖分生区细胞中的微管

用间接免疫荧光法在荧光显微镜下可以观察到植物根尖细胞分裂周期中微管捧列的变化.本文通过这一免疫细胞化学的方法研究了洋葱根尖分生区细胞中的微管周期.结果表明洋葱根尖细胞周期中有四种微管排列的方式,且这四种排列方式是一个循序渐进变化的过程,即周质微管、早前期带微管、纺锤体微管和成膜体微管.这四种微管的出现与细胞分裂周期密切相关,构成了典型的细胞微管周期.     

脾虚证动物模型复健后十二指肠粘膜上皮细胞的动力变化

脾虚证动物经四君子汤复健后的十二指肠粘膜上皮细胞动力学的变化,与大黄致脾虚动物相比,呈鲜明对照。隐窝细胞分裂速度及细胞周期变慢,绒毛长度增加,形成细胞寿命延长。初步认为绒毛显著变长,利于增强小肠吸收功能,似乎是脾虚动物达到复健的重要因素之一。     

Cyclopamine对牛脂肪间充质干细胞细胞周期相关基因表达的影响

间充质干细胞具有自我更新和多向分化的能力,能在体外诱导成脂细胞、软骨细胞、骨细胞和肌细胞等.脂肪间充质干细胞(adipose mesenchymal stem cells,AMSCs)取材方便,来源较为丰富.本试验利用hedgehog信号通路的特异性抑制剂cyclopamine(CPA)对牛AMSCs的细胞分裂周期机制做了初步探讨.先用血清饥饿法使细胞处于G0/G1期,再经CPA处理,检测不同处理时间的细胞周期蛋白cyclinA、cyclinD和cyclinE的表达情况.经CPA处理的细胞,其周期蛋白cyclin A、cyclin D和cyclin E的表达水平显著低于对照组.表明,CPA通过抑制细胞周期蛋白基因的表达从而抑制了细胞的增殖.     

抑癌基因p16的研究进展

p16是近年来确定的多肿瘤抑制基因，它对细胞周期的负调控作用尤为重要，其突变或缺失会导致对细胞分裂调控的丧失和紊乱，导致细胞的异常增殖，最终形成肿瘤。该基因在各种肿瘤中的作用，目前正越来越引起人们的重视。     

二氧化硫对菘蓝根尖生长及细胞分裂的影响

以二倍体菘蓝为材料,研究二氧化硫对菘蓝根尖生长及细胞分裂的影响,结果表明:低浓度二氧化硫对菘蓝根尖生长及细胞分裂没有影响;高浓度的(1.5～2.5mmol/L)二氧化硫能够抑制菘蓝根尖的生长,同时使细胞有丝分裂指数、发芽率下降,并诱发细胞分裂异常,且呈明显的时间效应和剂量效应。     

禾谷炭疽菌中3个典型Septin的生物信息学分析

septin是一个广泛存在于除植物以外的真菌、线虫、果蝇等生物的基因家族,其功能主要涉及细胞分裂、细胞极化、细胞内物质运输、囊泡运输和胞膜重构、细胞周期的调控与凋亡等生物学过程。前期研究发现,禾谷炭疽菌中含有三个典型的septin,分别为GLRG_07643、GLRG_03149、GLRG_07232.基于此,本研究从理化性质、二级结构以及亚细胞定位等方面对septin进行生物信息学分析,以期为深入开展septin基因的定位、表达以及功能域等方面研究,同时,为进一步实现以控制病原菌细胞分裂功能新的药物靶标的开发提供有力的理论支撑.     

亚历山大藻细胞周期中毒素含量及组成的变化

用细胞同步化培养的方法,研究了塔玛亚历山大藻A.tamarense CI01株不同细胞周期细胞中毒素含量与组成的变化.结果表明,A.tamarense CI01细胞分裂主要发生在早上6时至8时,细胞密度在6 h内增加近一倍.细胞内毒素合成主要在细胞周期的G1期,由光诱导合成,毒素含量(fmol/cell,Qt)在光周期前8 h内从17 fmol/cell增加到分裂前的24 fmol/cell,而在其它细胞周期细胞内毒素含量稳定,基本不合成毒素.比较分析单细胞C1,2毒素和GTX2,3毒素合成时间及速率,推测在A.tamarense CI01中细胞首先合成GTX2,3,在其它修饰酶的作用下再转化为C1,2.     

Cu~(2+)对冬小麦细胞分裂及叶绿体超微结构的影响

本文以冬小麦为实验材料,经不同浓度铜溶液处理,研究了铜肋迫对幼苗生长、细胞分裂及叶绿体超微结构的影响。实验结果表明:不同浓度的铜溶液可不同程度地抑制幼苗的生长,并导致根尖细胞有丝分裂指数的下降。较高浓度的氯化铜溶液(50ppm)处理可破坏叶绿体的膜系统,使叶绿体基粒垛迭减少,排列不规则。     

Regulation of the bacterial cell cycle by an integrated genetic circuit

How bacteria regulate cell cycle progression at a molecular level is a fundamental but poorly understood problem. In Caulobacter crescentus, two-component signal transduction proteins are crucial for cell cycle regulation, but the connectivity of regulators involved has remained elusive and key factors are unidentified. Here we identify ChpT, an essential histidine phosphotransferase that controls the activity of CtrA, the master cell cycle regulator. We show that the essential histidine kinase CckA initiates two phosphorelays, each requiring ChpT, which lead to the phosphorylation and stabilization of CtrA. Downregulation of CckA activity therefore results in the dephosphorylation and degradation of CtrA, which in turn allow the initiation of DNA replication. Furthermore, we show that CtrA triggers its own destruction by promoting cell division and inducing synthesis of the essential regulator DivK, which feeds back to downregulate CckA immediately before S phase. Our results define a single integrated circuit whose components and connectivity can account for the cell cycle oscillations of CtrA in Caulobacter.     

Caulobacter flagellin mRNA segregates asymmetrically at cell division

Molecular processes which promote the spatial localization of subcellular components are fundamental to cell development and differentiation. At various stages in development unequal segregation of molecular information must occur to result in the differentiated characteristics which distinguish cell progeny. Biological attributes of the dimorphic bacterium, Caulobacter crescentus, provide an experimental system permitting examination of the generation of asymmetry at the molecular level. When a Caulobacter cell divides, two different daughter cells are produced--a motile swarmer cell with a polar flagellum and a non-motile cell with a static appendage referred to as a stalk. The two cell types are distinct with respect to surface morphology, developmental potential, protein composition and biosynthetic capabilities. One of the more conspicuous manifestations of asymmetric expression of macromolecules in this system, the flagellum, has been studied extensively. We have cloned the flagellin genes of Caulobacter and report here the use of these sequences as probes to demonstrate that (1) the level of flagellin mRNA is regulated during the cell cycle in a pattern coincident with flagellum polypeptide synthesis and (2) flagellin mRNA synthesized before cell division is segregated with progeny swarmer cells. This provides molecular evidence of specific partitioning of an mRNA at the time of cell division.     

用蚕豆(Vicia faba)根尖细胞的微核细胞率作为环境监测指标的探讨

本试验表明,蚕豆根尖细胞的微核细胞率与水环境中污染物的质与量有密切的关系,对污染物有一定的敏感性,可作为监测环境污染状况的细胞学指标.同时,利用蚕豆根尖细胞的微核技术来监测环境污染状况还具有实验周期短、方法简便、鉴定可靠,又能反映出污染物对生物体的遗传伤害等优点.但要注意处理时温度对微核细胞率的影响.     

A mouse knock-in model exposes sequential proteolytic pathways that regulate p27Kip1 in G1 and S phase

The protein p27Kip1 is an inhibitor of cell division. An increase in p27 causes proliferating cells to exit from the cell cycle, and a decrease in p27 is necessary for quiescent cells to resume division. Abnormally low amounts of p27 are associated with pathological states of excessive cell proliferation, especially cancers. In normal and tumour cells, p27 is regulated primarily at the level of translation and protein turnover. Phosphorylation of p27 on threonine 187 (T187) by cyclin-dependent kinase 2 (Cdk2) is thought to initiate the major pathway for p27 proteolysis. To critically test the importance of this pathway in vivo, we replaced the murine p27 gene with one that encoded alanine instead of threonine at position 187 (p27T187A). Here we show that cells expressing p27T187A were unable to downregulate p27 during the S and G2 phases of the cell cycle, but that this had a surprisingly modest effect on cell proliferation both in vitro and in vivo. Our efforts to explain this unexpected result led to the discovery of a second proteolytic pathway for controlling p27, one that is activated by mitogens and degrades p27 exclusively during G1.     

离体培养中黄槐叶片细胞脱分化的细胞学观察

黄槐叶片外植体接种于 MS+2,4-D 1 ppm+NAA 1 ppm+6-BA 2 ppm 培养基上,外植体的维管薄壁细胞及维管束鞘细胞先脱分化启动,而后栅栏组织细胞脱分化,进而形成愈伤组织.细胞脱分化启动时有两个明显特点:一是细胞质变浓,核相对变大,核仁明显;二是细胞开始积累淀粉.细胞脱分化过程中细胞分裂方式以无丝分裂为主,有丝分裂较少.通过观察无丝分裂的主要过程,可以认为它是典型的劈裂式无丝分裂.文中还对两种细胞分裂方式在愈伤组织形成中的意义进行了讨论.     

Cell cycle oscillation of phosphatase inhibitor-2 in rat fibroblasts coincident with p34cdc2 restriction

Attention has focused on the regulation of the eucaryotic cell division cycle since the protein kinase p34cdc2 was identified as a key enzyme in mitotic induction. The level of this kinase remains constant throughout the cell cycle but its activity alters, particularly before M phase. Although the factors regulating cdc2 activity are still unknown, there is increasing evidence that it is influenced by p34cdc2 dephosphorylation. Protein phosphatase inhibitor-2 (I2) is a specific inhibitor of phosphatase type-1, which with type-2A is one of the two principal Ser(P) and Thr(P) phosphatases. Here we show that the level of I2, assayed by immunofluorescence staining, activity measurements, western immunoblotting and metabolic labelling, oscillates during the cell cycle in rat fibroblasts, peaking at S phase and mitosis. Moreover, when we inhibited I2 in vivo by microinjection of anti-I2 antibodies in S-phase cells, the pseudo-mitotic cellular response to injected p34cdc2 was restored, indicating that I2 might have a role in the modulation of p34cdc2 activity.     

Epistatic gene interactions in the control of division in fission yeast

THERE is currently much interest in the mechanism which controls the timing of cell division. Certain features of the control have been found to be common to a variety of eukaryotes. In particular, the importance of cell size as a parameter affecting cell cycle progress has been reported for mammalian cells(1,2) and for several single-celled eukaryotes(3-6). Another feature common to several systems is that growth conditions have a direct effect on the timing of division cycle events(7-9), and on cell size(9,10). In the fission yeast Schizosaccharomyces pombe, both cell size(6) and nutritional conditions(9) have been shown to affect cycle kinetics. The organism has been used extensively as a model eukaryotic system, largely because of the ease of measuring cell size and because division occurs by binary fission(11). More recently, its genetic tractability has led to the isolation of cell division cycle (cdc) mutants(12), and also of wee mutants altered in the control coordinating growth with the division cycle(13-15). The existence of such control mutants allows a more direct approach to the investigation of the molecular basis of division control, in contrast to the indirect methods used in other systems(4,16-18). wee mutants are so far unique to S. pombe. The most conspicuous property of wee mutants is their reduced cell size(13,14). Analysis of these mutants(15,19) and other evidence(9) has shown that control over cell division timing normally acts at entry to mitosis. As the function of a number of cdc genes is specifically required for mitosis(12), interactions between wee and cdc mutants which affect mitosis might be expected. I report here that the mitotic defect caused by a defective cdc25 allele is suppressed in wee mutants. Suppression by wee1 mutants is almost complete, while the wee2.1 mutation is a less effective suppressor. The significance of these findings for genetic models of the control of mitosis is considered.     

用蚕豆根尖细胞微核技术研究发芽马铃薯的遗传毒理作用

为探讨发芽马铃薯的遗传毒理作用,用蚕豆根尖细胞微核监测技术对发芽马薯浸渍液的微核率进行了测定,结果表明：发芽马铃薯浸渍液的质量浓度为25g/L,50g/L,100g/L,200g/L时,蚕豆根尖细胞微核率分别为17．21‰,23．53‰,29．55‰,39．40‰,与阴性对照组相比存在显著性差异,相应浓度的未发芽马铃薯浸渍液对蚕豆根尖细胞微核率的形成与阴性对照组相比无显著性差异,实验结果提示发芽马铃薯浸渍液能显著诱发蚕豆根尖细胞微核的形成,利用蚕豆根尖细胞微核技术可以监测发芽马铃薯的遗传毒理作用。     

Genome-wide survey of protein kinases required for cell cycle progression

Cycles of protein phosphorylation are fundamental in regulating the progression of the eukaryotic cell through its division cycle. Here we test the complement of Drosophila protein kinases (kinome) for cell cycle functions after gene silencing by RNA-mediated interference. We observed cell cycle dysfunction upon downregulation of 80 out of 228 protein kinases, including most kinases that are known to regulate the division cycle. We find new enzymes with cell cycle functions; some of these have family members already known to phosphorylate microtubules, actin or their associated proteins. Additionally, depletion of several signalling kinases leads to specific mitotic aberrations, suggesting novel roles for familiar enzymes. The survey reveals the inter-digitation of systems that monitor cellular physiology, cell size, cellular stress and signalling processes with the basic cell cycle regulatory machinery.