A polo-like kinase modulates cytokinesis and flagella biogenesis in Giardia lamblia

Background: Polo-like kinases (PLKs) are conserved serine/threonine kinases that regulate the cell cycle. To date, the role of Giardia lamblia PLK (GlPLK) in cells has not been studied. Here, we report our investigation on the function of GlPLK to provide insight into the role of this PKL in Giardia cell division, especially during cytokinesis and flagella formation. Methods: To assess the function of GIPLK, Giardia trophozoites were treated with the PLK-specific inhibitor GW843286X (GW). Using a putative open reading frame for the PLK identified in the Giardia genomic database, we generated a transgenic Giardia expressing hemagglutinin (HA)-tagged GlPLK and used this transgenic for immunofluorescence assays (IFAs). GlPLK expression was knocked down using an anti-glplk morpholino to observe its effect on the number of nuclei number and length of flagella. Giardia cells ectopically expressing truncated GlPLKs, kinase domain + linker (GlPLK-KDL) or polo-box domains (GlPLK-PBD) were constructed for IFAs. Mutant GlPLKs at Lys51, Thr179 and Thr183 were generated by site-directed mutagenesis and then used for the kinase assay. To elucidate the role of phosphorylated GlPLK, the phosphorylation residues were mutated and expressed in Giardia trophozoites RESULTS: After incubating trophozoites with 5 μM GW, the percentage of cells with > 4 nuclei and longer caudal and anterior flagella increased. IFAs indicated that GlPLK was localized to basal bodies and flagella and was present at mitotic spindles in dividing cells. Morpholino-mediated GlPLK knockdown resulted in the same phenotypes as those observed in GW-treated cells. In contrast to Giardia expressing GlPLK-PBD, Giardia expressing GlPLK-KDL was defective in terms of GIPLK localization to mitotic spindles and had altered localization of the basal bodies in dividing cells. Kinase assays using mutant recombinant GlPLKs indicated that mutation at Lys51 or at both Thr179 and Thr183 resulted in loss of kinase activity. Giardia expressing these mutant GlPLKs also demonstrated defects in cell growth, cytokinesis and flagella formation. Conclusions: These data indicate that GlPLK plays a role in Giardia cell division, especially during cytokinesis, and that it is also involved in flagella formation.ope

직무만족의 매개효과를 중심으로

학위논문(석사) -- 서울대학교대학원 : 행정대학원 공기업정책학과, 2021.8. 박순애.Techniques, which are recently emphasized from the perspective of human resource management, on the organizational commitment of public corporation employees through job satisfaction mediation: education and training, performance evaluation, position system, promotion system, and performance compensation. The purpose of this study was to analyze and determine whether the mediating effect of job satisfaction showed a difference according to gender. In other words, this study established a research model in which human resource management as an independent variable, job satisfaction as a parameter, and organizational commitment as a dependent variable, under this awareness of the problem, the indirect effect of human resource management on organizational commitment through job satisfaction as a medium. The effect of phosphorus was tested and the mediating effect mediated by gender was analyzed. The subject of this study is public corporation employees working for K Corporation. The survey results of 5,418 people were analyzed. The analysis results and implications are summarized as follows. This study reveals that human resource management not only has a direct effect on organizational commitment, but also has a significant indirect effect through job satisfaction as a mediator, thus showing a big direction that human resource management should be carried out by enhancing the job satisfaction of employees of public enterprises. presented. In addition, by verifying how each human resource management technique has different effects on organizational commitment through mediation of organizational commitment and job satisfaction of employees of public companies, and verifying the mediating effect adjusted to how these effects vary according to gender, the following results are obtained. Specific practical implications were obtained. First, as shown in the study results, the fact that human resource management has a positive (+) effect on job satisfaction and organizational commitment in general is considered to be sufficient incentive to utilize the human resource management system at the management strategic level of public corporations. can Second, it is also significant in that the strength of the relationship that human resource management mediates job satisfaction and leads to organizational commitment varies according to each human resource management technique. It was found that education and training system and personnel evaluation system had more influence on job satisfaction and organizational commitment than other factors. This proves that two factors among the human resource management implemented by public corporations are effective in increasing employee job satisfaction and organizational commitment compared to other factors. Third, it was found that performance compensation items did not have a satisfactory effect on organizational commitment directly among human resource management activities. This is judged to suggest that there is a potential difference of opinion or dissatisfaction with respect to performance compensation. Fourth, it can be seen that the effect of position management on organizational commitment by mediating job satisfaction is greater for women than for men. Through this, it can be seen that it is necessary to establish a balanced position assignment model that can increase the job satisfaction and organizational commitment of female employees in public corporations. If the management of position management of female employees in public corporations is strengthened, this will lead to job satisfaction of female employees. If the value is high, the effect of strengthening organizational commitment will be great. However, it is necessary to keep in mind the following limitations in interpreting the results of this study. Although this study was conducted using the results of a survey conducted by K Corporation, detailed measurement could not be carried out because the measurement items were simply designed for each variable. In future research, it will be possible to develop a scale through detailed questionnaire composition for each variable for more detailed measurement. In addition, the survey on human resource management is a survey data regularly conducted by K Corporation, and there is a possibility that employees will usually give a high score when answering the questionnaire. In addition, it would be more useful if a longitudinal study was added to examine the link between human resource management, job satisfaction, and organizational commitment. In addition, there is a limitation in that the problem of potential endogeneity was not properly controlled in that the analysis was performed using cross-sectional data observed at a specific time point. Therefore, in future studies, it is necessary to alleviate the problem of potential endogeneity by discovering appropriate instrumental variables or conducting analysis using panel data.본 연구는 최근 인적자원관리의 관점에서 강조되고 있는 교육훈련, 성과평가, 보직제도, 승진제도, 성과보상의 다섯 가지 인적자원관리 기법이 직무만족도의 매개를 통해 공기업 종사자의 조직몰입에 미치는 영향을 분석하고, 이러한 직무만족도의 매개효과가 성별에 따라 차이를 보이는지 확인하고자 하였다. 즉 본 연구는 이러한 문제의식 하에 인적자원관리를 독립변수로, 직무만족도를 매개변수로, 조직몰입을 종속변수로 하는 연구모형을 설정하여 인적자원관리가 직무만족도를 매개로 하여 조직몰입에 미치는 간접적인 영향을 검정하였고 성별에 의한 조절된 매개효과를 분석하였다. 연구대상은 K공사에 근무하는 공기업 종사자이며 총 5,418명의 설문조사 결과를 분석하였다. 분석 결과와 시사점을 요약하면 다음과 같다. 본 연구는 인적자원관리가 조직몰입에 직접적인 영향을 미칠 뿐만 아니라 직무만족도를 매개를 통해 간접적으로 미치는 영향도 상당하다는 점을 밝힘으로써 공기업 종사자들의 직무만족도 제고를 통한 인적자원관리가 이루어져야 한다는 큰 방향을 제시하였다. 또한 각각의 인적자원관리 기법별로 공기업 종사자의 조직몰입 및 직무만족의 매개를 통해 조직몰입에 미치는 영향이 어떻게 다른지 확인하고, 이러한 효과가 성별에 따라 어떻게 달라지는지 조절된 매개효과를 검증함으로써 다음과 같이 구체적인 실무적 시사점을 얻을 수 있었다. 첫째, 연구결과에 나타났듯이 전체적으로 볼 때 인적자원관리가 직무만족과 조직몰입에 정(+)의 영향을 주는 것으로 나타난 점은 공기업의 경영 전략적 차원에서도 인적자원관리 제도를 활용할 유인이 충분하다고 볼 수 있다. 둘째, 인적자원관리가 직무만족을 매개하여 조직몰입으로 이어지는 관계의 강도가 각각의 인적자원관리 기법에 따라 달라진다는 점에서도 의미하는 바가 크다. 교육훈련제도와 인사평가 제도 항목이 다른 요소들에 비해 직무만족이나 조직몰입에 더 많은 영향을 주는 것으로 나타났다. 이는 공기업에서 시행하고 있는 인적자원관리 가운데 두 가지 요소가 타 요소에 비해 종업원들의 직무만족도를 높이고 조직몰입도를 향상시키는데 유효함을 입증한 것이라 하겠다. 셋째, 인적자원관리활동 중 성과보상 항목이 직접적으로 조직 몰입에 대해 만족할 만한 영향을 주지 못한 것으로 나타났다. 이는 성과보상에 대한 견해 차이나 불만이 잠재되어 있음을 시사하는 것으로 판단된다. 넷째, 보직관리가 직무만족도를 매개하여 조직몰입도에 미치는 영향이 남성보다 여성에게 크다는 것을 알 수 있다. 이를 통해 공기업의 여성 종사자들의 직무만족도와 조직몰입도를 높일 수 있는 균형적인 보직배치 모델 구축이 필요하다는 점을 알 수 있고 공기업의 여성 종사들의 보직관리에 대한 관리를 강화한다면 이는 여성 종사자들의 직무 만족도를 높게 해주면 조직몰입도를 강화하는 효과가 클 것이다. 다만, 본 연구의 결과를 해석함에 있어서 다음과 같은 한계에 유념할 필요가 있다. 본 연구는 K공사에서 실시하는 설문조사 결과를 활용하여 진행하였지만 변수별로 측정문항이 단순하게 설계되어 있어 세밀한 측정이 진행되지 못하였다. 향후 연구에서는 보다 세밀한 측정을 위해 각 변수별로 세밀한 설문 구성을 통해 척도 개발을 할 수 있을 것이다. 또한 인적자원관리에 대한 설문조사가 K공사에서 정기적으로 실시하는 설문자료로 이는 직원들이 답변시 통상 높은 점수를 부여할 개연성이 있다. 더불어 인적자원관리와 직무만족도, 조직몰입 간 연계를 살펴보기 위해서는 종단적 연구가 추가된다면 더 유용하겠지만 본 연구에서는 횡단적인 연구방법을 실시하였기 때문에 시간의 흐름에 따른 변화를 반영하지 못하였다. 또한 특정 시점에 관측된 횡단자료를 사용하여 분석이 이루어졌다는 점에서 잠재적 내생성의 문제를 적절히 통제하지 못하였다는 한계가 있다. 따라서 추후 연구에서는 적절한 도구변수를 발굴하거나 패널자료를 사용하여 분석을 진행하여 잠재적 내생성의 문제를 완화시킬 필요가 있다.제 1 장 서 론 1 제 1 절 연구의 필요성 및 목적1 1. 연구의 배경 1 2. 연구의 목적 3 제 2 절 연구대상과 연구방법 7 1. 연구대상 및 범위 7 2. 연구의 방법 8 제 2 장 이론적 논의와 선행연구 검토 9 제 1 절 인적자원관리 9 1. 인적자원관리 개념 9 2. 인적자원관리의 구성요소 11 3. 인적자원관리의 중요성 12 제 2 절 조직몰입 15 1. 조직몰입 개념 16 2. 조직몰입의 중요성 18 제 3 절 직무만족 19 1. 직무만족 개념 19 2. 직무만족의 중요성22 제 4 절 선행연구 검토 및 가설설정 23 1. 인적자원관리와 직무만족 23 2. 인적자원관리와 조직몰입 25 3. 직무만족과 조직몰입 27 4. 인적자원관리와 조직몰입간의 직무만족의 매개효과 30 5. 성별차이 : 조절된 매개효과 31 제 5 절 K공사의 인적자원관리 32 1. 교육훈련32 2. 인사평가34 3. 보직관리 36 4. 승진제도 37 5. 성과보상 38 제 3 장 연구 설계 39 제 1 절 연구 모형 39 제 2 절 변수의 정의 및 측정도구 40 1. 독립변수 40 2. 종속변수 41 3. 매개변수 41 4. 조절변수 41 4. 통제변수 42 제 3 절 자료 수집 및 분석방법 42 제 4 장 연구결과 43 제 1 절 연구대상의 특성 43 제 2 절 기술통계 45 제 3 절 상관관계 46 제 4 절 연구대상의 일반적 특성에 따른 차이 47 1. 연령 47 2. 성별 49 3. 소속 50 4. 직군 52 5. 직급 54 제 5 절 인적자원관리,직무몰입, 직무만족 간 영향관계 56 1. 인적자원관리에 대한 인식이 직무만족도에 미치는 영향 56 2. 인적자원관리와 직무만족도가 조직몰입도에 미치는 영향 57 3. 직무만족의 매개효과 59 4. 성별의 조절효과 및 조절된 매개효과 61 제 5장 결론 65 제 1 절 연구결과 요약 및 시사점 65 1. 연구결과 요약 67 2. 연구의 의의 및 시사점 67 제 2 절 연구의 한계 및 향후 연구과제 70 참고문헌 73석

Characterization of microtubule-binding and dimerizatino activity of Giardia lamblia end-binding 1 protein

End-binding 1 (EB1) proteins are evolutionarily conserved components of microtubule (MT) plus-end tracking protein that regulate MT dynamics. Giardia lamblia, with two nuclei and cytoskeletal structures, requires accurate MT distribution for division. In this study, we show that a single EB1 homolog gene of G. lamblia regulates MT dynamics in mitosis. The haemagglutinin-tagged G. lamblia EB1 (GlEB1) localizes to the nuclear envelopes and median bodies, and is transiently present in mitotic spindles of dividing cells. Knockdown of GlEB1 expression using the morpholinos-based anti-EB1 oligonucleotides, resulted in a significant defect in mitosis of Giardia trophozoites. The MT-binding assays using recombinant GlEB1 (rGlEB1) proteins demonstrated that rGlEB1102-238, but not rGlEB11-184, maintains an MT-binding ability comparable with that of the full length protein, rGlEB11-238. Size exclusion chromatography showed that rGlEB1 is present as a dimer formed by its C-terminal domain and a disulfide bond. In vitro-mutagenesis of GlEB1 indicated that an intermolecular disulfide bond is made between cysteine #13 of the two monomers. Complementation assay using the BIM1 knockout mutant yeast, the yeast homolog of mammalian EB1, indicated that expression of the C13S mutant GlEB1 protein cannot rescue the mitotic defect of the BIM1 mutant yeast. These results suggest that dimerization of GlEB1 via the 13th cysteine residues plays a role during mitosis in Giardia.ope

Role of gamma-giardin in ventral disc formation of Giardia lamblia

BACKGROUND: Giardia lamblia, a protozoan pathogen causing diarrheal outbreaks, has characteristic cytoskeletal structures including eight flagella, a median body and a ventral disc. Gamma-giardin is a unique component protein of the cytoskeleton of this protozoan. RESULTS: Through comparative proteomic analysis between different stages of the cell cycle, G. lamblia γ-giardin (Glγ-giardin) was identified as an upregulated protein in the G2-phase. Increased Glγ-giardin expression in G2 was confirmed by western blot and real-time polymerase chain reaction analyses. Knockdown of this protein using a morpholino affected the formation of ventral discs, especially the microribbons of the discs, but exerted little effect on the binding ability of G. lamblia. The number of cells with four nuclei was increased in Glγ-giardin-knockdown cells. Expression of Glγ-giardin was decreased during encystation, in contrast with the G2-phase. CONCLUSIONS: Knockdown experiments demonstrated that Glγ-giardin is a component of the trilaminar structure of the ventral disc. Expression of Glγ-giardin is induced in the G2-phase prior to active cell division, whereas its expression decreases during encystation, a dormant stage of G. lamblia.ope

Kinesin-13, a Motor Protein, is Regulated by Polo-like Kinase in Giardia lamblia

Kinesin-13 (Kin-13), a depolymerizer of microtubule (MT), has been known to affect the length of Giardia. Giardia Kin-13 (GlKin-13) was localized to axoneme, flagellar tips, and centrosomes, where phosphorylated forms of Giardia polo-like kinase (GlPLK) were distributed. We observed the interaction between GlKin-13 and GlPLK via co-immunoprecipitation using transgenic Giardia cells expressing Myc-tagged GlKin-13, hemagglutinin-tagged GlPLK, and in vitro-synthesized GlKin-13 and GlPLK proteins. In vitro-synthesized GlPLK was demonstrated to auto-phosphorylate and phosphorylate GlKin-13 upon incubation with [γ-32P]ATP. Morpholino-mediated depletion of both GlKin-13 and GlPLK caused an extension of flagella and a decreased volume of median bodies in Giardia trophozoites. Our results suggest that GlPLK plays a pertinent role in formation of flagella and median bodies by modulating MT depolymerizing activity of GlKin-13.ope

Trichomonas vaginalis α-Actinin 2 Modulates Host Immune Responses by Inducing Tolerogenic Dendritic Cells via IL-10 Production from Regulatory T Cells

Trichomonas vaginalis is a pathogen that triggers severe immune responses in hosts. T. vaginalis α-actinin 2, Tvα-actinin 2, has been used to diagnose trichomoniasis. This study was undertaken to examine the role of Tvα-actinin 2 as an antigenic molecule to induce immune responses from humans. Western blot analysis using anti-Tvα-actinin 2 antibodies indicated its presence in the secreted proteins of T. vaginalis. ELISA was employed to measure cytokine production by vaginal epithelial cells, prostate cells, mouse dendritic cells (DCs), or T cells stimulated with T. vaginalis or Tvα-actinin 2 protein. Both T. vaginalis and rTvα-actinin 2 induced cytokine production from epithelial cell lines, including IL-10. Moreover, CD4+CD25- regulatory T cells (Treg cells) incubated with rTvα-actinin 2-treated DCs produced high levels of IL-10. These data indicate that Tvα-actinin 2 modulates immune responses via IL-10 production by Treg cells.ope

A Standard Public Order Treaty Carve-out as a Means for Balancing Regulatory Interests in International Investment Agreements

학위논문 (박사)-- 서울대학교 대학원 : 법학과, 2017. 2. 신희택.Since the start of the BIT program, the purpose of BITs was predominantly motivated by the sole objective of providing foreign investment protection mostly for the benefit of the capital-exporting, developed countries. The capital-importing, developing countries entered into BITs to attract foreign investment without a comprehensive awareness of the legal ramifications of BITs. However, the conclusion of FTAs with investment chapters such as the NAFTA expanded the narrowly conceived BIT goal of investment protection to include other purposes like investment promotion and liberalization. Prior to the NAFTA, carving out policy space in international investment agreements (IIAs) was not a real concern until the NAFTA experience demonstrated that investor-State arbitrations could be initiated not only against the developing States, but also against the developed States. Moreover, the ICSID cases against Argentina have been instrumental in bringing attention to the need of host States to exercise regulatory power. These experiences helped to create an understanding for both the developed and developing States that a significant legal consequence of concluding IIAs is that their sovereign right to regulate various aspects of public interest might result in a breach of the IIA. The objective of this research is to fill a meaningful gap in international investment law to enable States to better exercise their sovereign right to regulate by using the public order clause in the non-precluded measures provision of the U.S.-Argentina BIT as a starting point. The questions asked in this study include whether a public order carve-out for public interest matters is emerging and, if so, whether the public order carve-out can equip host States with the flexibility needed to exercise their regulatory authority. This research makes the discovery that the concept of public order is undefined in international investment law making it difficult for investment tribunals to interpret the public order carve-out in a consistent and predictable manner. On one level, the notion of the right to regulate is being incorporated in the most current versions of IIAs without an appreciation of how it should apply in international investment law despite the lessons exemplified in the ICSID arbitrations against Argentina. On another level, BITs have been designed to usually only contain substantive obligations. Although the recent trend of IIAs is to include some variation of a general exceptions provision to limit the scope of the substantive obligations, the practice remains largely inconsistent and borrowed from the WTO/GATS jurisprudence. However, this research concludes that the inclusion of a standard public order carve-out specifically aimed at preserving the regulatory space of States should become a fixed feature of future investment treaties to better address the growing aggregate community interests of IIA stakeholders. This ultimately requires that the base values and concerns of the participants in international investment law be evaluated.1. Introduction 1 1.1. Trends of International Investment Agreements 1 1.1.A. Changes in BIT/IIA Perspective 1 1.1.B. Aggregate Community Interests of the IIA Stakeholders 16 1.2. Problem Identification 54 2. Preserving Regulatory Space through the Public Order Carve-out Provision in IIAs 62 2.1. The Multiple Meanings of "Public Order" 67 2.1.A. The National Law Concept of Public Order 65 2.1.B. The International Law Concept of Public Order 72 2.2. The State Concept of Public Order in the Context of International Organizations 78 2.2.A. Public Order in the European Union 78 2.2.B. Public Order in the WTO/GATS 86 2.2.C. Public Order in Human Rights Conventions 92 2.3. The Customary International Law Source of the Public Order Concept 105 2.4. Concluding Remarks 117 3. States' Treatment of the Public Order Carve-out in IIAs 121 3.1. Textual Transformation of the Public Order Carve-out in IIAs 121 3.1.A. Prior to the 1980s: Public Order Provisions in FCNs 121 3.1.B. Prior to the 2000s: Public Order Provisions in BITs 125 3.1.C. Current Trends: FTA Investment Chapters with Regulatory Space Carve-outs 131 3.2. Other Methods of Preserving Regulatory Space in IIAs 154 3.2.A. Legitimate Public Welfare Objectives 154 3.2.B. Reservations 164 3.2.C. National Security 170 3.2.D. Essential Security Interest 184 3.3. Concluding Remarks 190 4. Treatment of the Public Order Carve-out by the ICSID Tribunals in Cases Arising out of the U.S.-Argentina BIT 192 4.1. Fact Pattern of the Argentine ICSID Cases 192 4.2. The Argentine ICSID Tribunals' Standard of Review for the Public Order Carve-out 200 4.2.A. ILC Article 25 as Applied to the Public Order Carve-out 200 4.2.B. The Argentine ICSID Tribunals' Treatment of the NPM Treaty Provision 217 4.3. Concluding Remarks 237 5. A Standard Public Order Carve-out in IIAs to Balance Stakeholders' Interests 241 5.1. Proposal of a Sample Standard Public Order Carve-out 241 5.1.A. Evolving Object and Purpose of IIAs 243 5.1.B. Scope of a Standard Public Order Carve-out 251 5.1.C. Distinguishing the Standard Public Order Carve-out from the WTO/GATS-inspired General Exceptions Clause 257 5.1.D. The Public Order Carve-out is not a Lex Specialis Rule 271 5.1.E. Preserving Regulatory Space in the Absence of the Public Order Carve-out 281 5.2. The Legitimate Policy Objectives in the Proposed Standard Public Order Carve-out as Explained by Existing Cases 293 5.3. Concluding Remarks 299 6. Conclusion 301 참고문헌 308 국문초록 346Docto

Role of α-Actinin 2 in Cytoadherence and Cytotoxicity of Trichomonas vaginalis

Trichomonas vaginalis is a pathogen that triggers severe immune responses in hosts. T. vaginalis α-actinin 2 (Tvα-actinin 2) has been used to diagnose trichomoniasis. Tvα-actinin 2 was dissected into three parts; the N-terminal, central, and C-terminal portions of the protein (#1, #2, and #3, respectively). Western blot of these Tvα-actinin 2 proteins with pooled patients' sera indicated that #2 and #3, but not #1, reacted with those sera. Immunofluorescence assays of two different forms of T. vaginalis (trophozoites and amoeboid forms), using anti-Tvα-actinin 2 antibodies, showed localization of Tvα-actinin 2 close to the plasma membranes of the amoeboid form. Fractionation experiments indicated the presence of Tvα-actinin 2 in cytoplasmic, membrane, and secreted proteins of T. vaginalis. Binding of fluorescence-labeled Trichomonas to vaginal epithelial cells and prostate cells was decreased in the antibody blocking experiment using anti-Tvα-actinin 2 antibodies. Pretreatment of T. vaginalis with anti-rTvα-actinin 2 antibodies also resulted in reduction in its cytotoxicity. Flow cytometry, ligand-binding immunoblotting assay, and observation by fluorescence microscopy were used to detect the binding of recombinant Tvα-actinin 2 to human epithelial cell lines. Specifically, the truncated N-terminal portion of Tvα-actinin 2, Tvα-actinin 2 #1, was shown to bind directly to vaginal epithelial cells. These data suggest that α-actinin 2 is one of the virulence factors responsible for the pathogenesis of T. vaginalis by serving as an adhesin to the host cells.ope

Functional Identification of a Nuclear Localization Signal of MYB2 Protein in Giardia lamblia

MYB2 protein was identified as a transcription factor that showed encystation-induced expression in Giardia lamblia. Although nuclear import is essential for the functioning of a transcription factor, an evident nuclear localization signal (NLS) of G. lamblia MYB2 (GlMYB2) has not been defined. Based on putative GlMYB2 NLSs predicted by 2 programs, a series of plasmids expressing hemagglutinin (HA)-tagged GlMYB2 from the promoter of G. lamblia glutamate dehydrogenase were constructed and transfected into Giardia trophozoites. Immunofluorescence assays using anti-HA antibodies indicated that GlMYB2 amino acid sequence #507-#530 was required for the nuclear localization of GlMYB2, and this sequence was named as NLSGlMYB2. We further verified this finding by demonstrating the nuclear location of a protein obtained by the fusion of NLSGlMYB2 and G. lamblia glyceraldehyde 3-phosphate dehydrogenase, a non-nuclear protein. Our data on GlMYB2 will expand our understanding on NLSs functioning in G. lamblia.ope

RNA-sequencing Profiles of Cell Cycle-Related Genes Upregulated during the G2-Phase in Giardia lamblia

To identify the component(s) involved in cell cycle control in the protozoan Giardia lamblia, cells arrested at the G1/S- or G2-phase by treatment with nocodazole and aphidicolin were prepared from the synchronized cell cultures. RNA-sequencing analysis of the 2 stages of Giardia cell cycle identified several cell cycle genes that were up-regulated at the G2-phase. Transcriptome analysis of cells in 2 distinct cellcycle stages of G. lamblia confirmed previously reported components of cell cycle (PcnA, cyclin B, and CDK) and identified additional cellcycle components (NEKs, Mad2, spindle pole protein, and CDC14A). This result indicates that the cell cycle machinery operates in this protozoan, one of the earliest diverging eukaryotic lineages.ope