A double lock on sister chromatids by cohesin

In this issue of Molecular Cell, Farcas et al. (2011) demonstrate that intertwining between sister chromatids at metaphase is much more significant than previously thought and, remarkably, show that it depends on cohesin.     

Proteins: caught in the trap

Two independent groups have recently devised innovative methods using light to trap and manipulate particles as small as proteins.     

Cleavage of cohesin rings coordinates the separation of centrioles and chromatids

Cohesin pairs sister chromatids by forming a tripartite Scc1-Smc1-Smc3 ring around them. In mitosis, cohesin is removed from chromosome arms by the phosphorylation-dependent prophase pathway. Centromeric cohesin is protected by shugoshin 1 and protein phosphatase 2A (Sgo1-PP2A) and opened only in anaphase by separase-dependent cleavage of Scc1 (refs 4-6). Following chromosome segregation, centrioles loosen their tight orthogonal arrangement, which licenses later centrosome duplication in S phase. Although a role of separase in centriole disengagement has been reported, the molecular details of this process remain enigmatic. Here, we identify cohesin as a centriole-engagement factor. Both premature sister-chromatid separation and centriole disengagement are induced by ectopic activation of separase or depletion of Sgo1. These unscheduled events are suppressed by expression of non-cleavable Scc1 or inhibition of the prophase pathway. When endogenous Scc1 is replaced by artificially cleavable Scc1, the corresponding site-specific protease triggers centriole disengagement. Separation of centrioles can alternatively be induced by ectopic cleavage of an engineered Smc3. Thus, the chromosome and centrosome cycles exhibit extensive parallels and are coordinated with each other by dual use of the cohesin ring complex.     

Membrane fusion: caught in a trap

SNAREs are small coiled-coil proteins required for specific membrane fusion events in eukaryotic cells. Recent evidence points to the existence of an inhibitory class of SNAREs, i-SNAREs, which prevent incorrect fusions from occurring, adding a further layer of regulation to the process of membrane docking and fusion.     

Absence of mouse REC8 cohesin promotes synapsis of sister chromatids in meiosis

REC8 is a key component of the meiotic cohesin complex. During meiosis, cohesin is required for the establishment and maintenance of sister-chromatid cohesion, for the formation of the synaptonemal complex, and for recombination between homologous chromosomes. We show that REC8 has an essential role in mammalian meiosis, in that Rec8 null mice of both sexes have germ cell failure and are sterile. In the absence of REC8, early chromosome pairing events appear normal, but synapsis occurs in a novel fashion: between sister chromatids. This implies that a major role for REC8 in mammalian meiosis is to limit synapsis to between homologous chromosomes. In all other eukaryotic species studied to date, REC8 phenotypes have been restricted to meiosis. Unexpectedly, Rec8 null mice are born in sub-Mendelian frequencies and fail to thrive. These findings illuminate hitherto unknown REC8 functions in chromosome dynamics during mammalian meiosis and possibly in somatic development.     

Statistical indicators for insects caught in the development trap

The effects of climate change on insect phenologies are of current interest, but little attention has been given to the potential driver for population decline of increased mortality arising from the failure to reach a viable overwintering stage. We use numerical simulation modelling to show that increased mortality from a partial second generation results in the weakening of density dependence, whereas increasing mortality within the first generation does not. These general results are then compared with annual trap count data for the moth Agrotis segetum (Denis and Schiffermüller) in Denmark, a species which is in decline there because a partial second generation fails to survive the winter. Results from the moth data are consistent with those from the simulations, and we conclude that the development trap phenomenon can be detected by the application of simple statistical tests to time‐series data.     

Sister chromatid resolution: a cohesin releasing network and beyond

When chromosomes start to assemble in mitotic prophase, duplicated chromatids are not discernible within each chromosome. As condensation proceeds, they gradually show up, culminating in two rod-shaped structures apposed along their entire length within a metaphase chromosome. This process, known as sister chromatid resolution, is thought to be a prerequisite for rapid and synchronous separation of sister chromatids in anaphase. From a mechanistic point of view, the resolution process can be dissected into three distinct steps: (1) release of cohesin from chromosome arms; (2) formation of chromatid axes mediated by condensins; and (3) untanglement of inter-sister catenation catalyzed by topoisomerase II (topo II). In this review article, we summarize recent progress in our understanding the molecular mechanisms of sister chromatid resolution with a major focus on its first step, cohesin release. An emerging idea is that this seemingly simple step is regulated by an intricate network of positive and negative factors, including cohesin-binding proteins and mitotic kinases. Interestingly, some key factors responsible for cohesin release in early mitosis also play important roles in controlling cohesin functions during interphase. Finally, we discuss how the step of cohesin release might mechanistically be coordinated with the actions of condensins and topo II.     

Sister chromatids fail to separate during an induced endoreplication cycle in Drosophila embryos

When mitosis is bypassed, as in some cancer cells or in natural endocycles, sister chromosomes remain paired and produce four-stranded diplochromosomes or polytene chromosomes. Cyclin/Cdk1 inactivation blocks entry into mitosis and can reset G2 cells to G1, allowing another round of replication. Reciprocally, persistent expression of Cyclin A/Cdk1 or Cyclin E/Cdk2 blocks Drosophila endocycles. Inactivation of Cyclin A/Cdk1 by mutation or overexpression of the Cyclin/Cdk1 inhibitor, Roughex (Rux), converts the 16(th) embryonic mitotic cycle to an endocycle; however, we show that Rux expression fails to convert earlier cell cycles unless Cyclin E is also downregulated. Following induction of a Rux transgene in Cyclin E mutant embryos during G2 of cell cycle 14 (G2(14)), Cyclins A, B, and B3 disappeared and cells reentered S phase. This rereplication produced diplochromosomes that segregated abnormally at a subsequent mitosis. Thus, like the yeast CKIs Rum1 and Sic1, Drosophila Rux can reset G2 cells to G1. The observed cyclin destruction suggests that cell cycle resetting by Rux was associated with activation of the anaphase-promoting complex (APC), while the presence of diplochromosomes implies that this activation of APC outside of mitosis was not sufficient to trigger sister disjunction.     

A SIR-tain acetyl complex is caught by a sulfur trap

In this issue, Hawse et al. (2008) provide additional insight into the mechanistic properties of sirtuin enzymes by describing the structure of a thio-imidate in the active site of Thermatoga maritima Sir2, which strengthens the proposal that the enzyme directly couples NAD(+) and acetyllysine oxygen to form a versatile ADPR-peptidyl-imidate intermediate.     

RSC facilitates Rad59-dependent homologous recombination between sister chromatids by promoting cohesin loading at DNA double-strand breaks

Homologous recombination repairs DNA double-strand breaks by searching for, invading, and copying information from a homologous template, typically the homologous chromosome or sister chromatid. Tight wrapping of DNA around histone octamers, however, impedes access of repair proteins to DNA damage. To facilitate DNA repair, modifications of histones and energy-dependent remodeling of chromatin are required, but the precise mechanisms by which chromatin modification and remodeling enzymes contribute to homologous DNA repair are unknown. Here we have systematically assessed the role of budding yeast RSC (remodel structure of chromatin), an abundant, ATP-dependent chromatin-remodeling complex, in the cellular response to spontaneous and induced DNA damage. RSC physically interacts with the recombination protein Rad59 and functions in homologous recombination. Multiple recombination assays revealed that RSC is uniquely required for recombination between sister chromatids by virtue of its ability to recruit cohesin at DNA breaks and thereby promoting sister chromatid cohesion. This study provides molecular insights into how chromatin remodeling contributes to DNA repair and maintenance of chromatin fidelity in the face of DNA damage.     

Influence of trap surface on the numbers of insects caught in water traps in brassica crops

The contribution of the various surfaces of a water trap to the trap's overall effectiveness was studied in brassica crops by painting black different parts of fluorescent-yellow water-traps. Three pest species, Delia radicum (L.), D. platura (Meig.), and Meligethes aeneus (Fab.)/Meligethes viridescens (Fab.), together with blowflies and syrphids, were caught in large numbers. Each insect responded differently to the yellow/black traps. The numbers of insect caught indicated that the area of trap involved in capturing D. radicum was effectively twice the surface area of the water. Yellow traps for monitoring D. radicum populations could be made more selective by painting the inner wall black, as such traps caught similar numbers of D. radicum to all-yellow traps but 50% fewer D. platura, Meligethes sp. and blowflies, and 95% fewer syrphids.     

A protein caught in a kinetic trap: structures and stabilities of insulin disulfide isomers

Proinsulin contains six cysteines whose specific pairing (A6-A11, A7-B7, and A20-B19) is a defining feature of the insulin fold. Pairing information is contained within A and B domains as demonstrated by studies of insulin chain recombination. Two insulin isomers containing non-native disulfide bridges ([A7-A11,A6-B7,A20-B19] and [A6-A7,A11-B7,A20-B19]), previously prepared by directed chemical synthesis, are metastable and biologically active. Remarkably, the same two isomers are preferentially formed from native insulin or proinsulin following disulfide reassortment in guanidine hydrochloride. The absence of other disulfide isomers suggests that the observed species exhibit greater relative stability and/or kinetic accessibility. The structure of the first isomer ([A7-A11,A6-B7,A20-B19], insulin-swap) has been described [Hua, Q. X., Gozani, S. N., Chance, R. E., Hoffmann, J. A., Frank, B. H., and Weiss, M. A. (1995) Nat. Struct. Biol. 2, 129-138]. Here, we demonstrate that the second isomer (insulin-swap2) is less ordered than the first. Nativelike elements of structure are retained in the B chain, whereas the A chain is largely disordered. Thermodynamic studies of guanidine denaturation demonstrate the instability of the isomers relative to native insulin (DeltaDeltaG(u) > 3 kcal/mol). In contrast, insulin-like growth factor I (IGF-I) and the corresponding isomer IGF-swap, formed as alternative products of a bifurcating folding pathway, exhibit similar cooperative unfolding transitions. The insulin isomers are similar in structure and stability to two-disulfide analogues whose partial folds provide models of oxidative folding intermediates. Each exhibits a nativelike B chain and less-ordered A chain. This general asymmetry is consistent with a hierarchical disulfide pathway in which nascent structure in the B chain provides a template for folding of the A chain. Structures of metastable disulfide isomers provide probes of the topography of an energy landscape.     

A new meiosis-specific cohesin complex implicated in the cohesin code for homologous pairing

We identify a new mammalian cohesin subunit, RAD21-like protein (RAD21L), with sequence similarity to RAD21 and REC8. RAD21L localizes along axial elements in early meiotic prophase, in a manner that is spatiotemporally different to either REC8 or RAD21. Remarkably, RAD21L and REC8 have symmetrical, mutually exclusive localization on the not-yet-synapsed homologues, implying that the cohesin patterning could provide a code for homologue recognition. RAD21 transiently localizes to axial elements after the dissociation of RAD21L and REC8 in late pachytene, a period of recombination repair. Further, we show that the removal of cohesins and synaptonemal complex during late meiotic prophase is promoted by Polo-like kinase 1, which is similar to the mitotic prophase pathway.     

Comparison of pheromone trap design and lures for Spodoptera frugiperda in Togo and genetic characterization of moths caught

Fall armyworm, Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae), is a pest of grain and vegetable crops endemic to the Western Hemisphere that has recently become widespread in sub‐Saharan Africa and has appeared in India. An important tool for monitoring S. frugiperda in the USA is pheromone trapping, which would be of value for use with African populations. Field experiments were conducted in Togo (West Africa) to compare capture of male fall armyworm using three commercially available pheromone lures and three trap designs. The objectives were to identify optimum trap × lure combinations with respect to sensitivity, specificity, and cost. Almost 400 moths were captured during the experiment. Differences were found in the number of S. frugiperda moths captured in the various trap designs and with the three pheromone lures, and in the number of non‐target moths captured with each lure. The merits of each trap × lure combination are discussed with respect to use in Africa. A nearly equal number of COI‐CS (161) and COI‐RS (158) moths was captured with no differences found in COI marker proportions among traps or lures. However, the diagnostic rice strain marker Tpi was rarely found. Overall, the genetic characterization of the pheromone trap collections indicated a consistent distribution of genetic markers from 2016 to 2017, suggesting a population at or near equilibrium.     

Evidence for the uninemy of eukaryotic chromatids

Polytene chromosomes of Chironomus thummi were stretched to their rupture in a pronase solution. A 176±26-fold elongation was achieved. The DNA compaction ratio, defined as the ratio of DNA length in a haploid set (85±5 mm) to the length of a polytene chromosome set (520±40 m), was 164±22. Closeness of these two values demonstrates the uninemy of the chromatids of Chironomus chromosomes. The effect of ethidium bromide on the elastic properties of chromosomes prestretched in a pronase solution and the lengthening of these chromosomes after ethidium staining suggest that DNA molecules are double-stranded and supercoiled to the moment of the chromosome rupture. It is concluded that a Chironomus chromatid consists of a single DNA molecule (or of a single chain of linked DNA molecules) both ends of which are located in the telomeres.To the memory of Prof. Vera V. Khvostova