• Reproductive and Developmental Biology
• /
• 제39권4호
• /
• pp.105-115
• /
• 2015

Toll-like receptor 7 (TLR7) is critical for the triggering of innate immune response by recognizing the conserved molecular patterns of single-stranded RNA (ssRNA) viruses and mediated antigenic adaptive immunity. To understand how TLR7 distinguish pathogen-derived molecular patterns from the host self, it is essential to be able to identify TLR7 receptor interaction interfaces, such as active sites or R848-agonist binding sites. The functional interfaces of TLR7 can serve as targets for structure-based drug design in studying the TLR7 receptor's structure-function relationship. In contrast to mammalian TLR7, chicken TLR7 (chTLR7) is unknown for its important biological function. Therefore, it has been targeted to mediate contrasting evolutionary patterns of positive selection into non-synonymous SNPs across eleven species using TLR7 conservation patterns (evolutionary conserved and class-specific trace residues), where protein sequence differences to the TLR7 receptors of interest record mutation that have passed positive section across the species. In this study, we characterized the Lys609 residue on chTLR7-ECD homodimer interfaces to reflect the current tendency of evolving positive selection to be transfer into a stabilization direction of the R848-agonist/chTLR7-ECDs complex under the phylogenetically variable position across species and we suggest a potential indicator for contrasting evolutionary patterns of both the species TLR-ECDs.

• Molecules and Cells
• /
• 제28권1호
• /
• pp.57-65
• /
• 2009

CDC48 is a member of the AAA ATPase superfamily. Yeast CDC48 and its mammalian homolog p97 are implicated in diverse cellular processes, including mitosis, membrane fusion, and ubiquitin-dependent protein degradation. However, the cellular functions of plant CDC48 proteins are largely unknown. In the present study, we performed virus-induced gene silencing (VIGS) screening and found that silencing of a gene encoding a tobacco CDC48 homolog, NgCDC48, resulted in severe abnormalities in leaf and shoot development in tobacco. Furthermore, transgenic tobacco plants (35S:anti-NgCDC48), in which the NgCDC48 gene was suppressed using the antisense RNA method, exhibited severely aberrant development of both vegetative and reproductive organs, resulting in arrested shoot and leaf growth and sterile flowers. Approximately 57-83% of 35S:anti-NgCDC48 plants failed to develop mature organs and died at early stage of development. Scanning electron microscopy showed that both adaxial and abaxial epidermal pavement cells in antisense transgenic leaves were significantly smaller and more numerous than those in wild type leaves. These results indicate that NgCDC48 is critically involved in cell growth and development of tobacco plants. An in vivo targeting experiment revealed that NgCDC48 resides in the endoplasmic reticulum (ER) in tobacco protoplasts. We consider the tantalizing possibility that CDC48-mediated degradation of an as-yet unidentified protein(s) in the ER might be a critical step for cell growth and expansion in tobacco leaves.

• Genomics & Informatics
• /
• 제3권4호
• /
• pp.142-148
• /
• 2005

The immune response-related genes have been suggested to be the most favorable genes for positive selection during evolution. Comparing the entire DNA sequence of chimpanzee chromosome 22 (PTR22) with human chromosome 21 (HSA21), we have identified 15 orthologs having indel in their coding sequences. Among them, interferon-${\alpha}$ receptor-1 gene (IFNAR1), an immuneresponse-related gene, is subjected to comparative genomic analysis. Chimpanzee IFNAR1 showed the same genomic structure as human IFNAR1 (11 exons and 10 introns) except the 3 bp insertion in exon 4. The sequence alignment of IFNAR1 coding sequence indicated that 'ISPP' amino acid sequence motif is highly conserved in chimpanzee and other animals including mouse and chicken. However, the human IFNAR1 shows that one proline residue is missing in the sequence motif. The homology modeling of the IFNAR1 structures suggests that the proline deletion in human IFNAR1 leads to the formation of the following ${\alpha}$-helix, whereas two sequential prolines in chimpanzee IFNAR1 inhibit it. As a result, human IFNAR1 may adopt a characteristic structure distinct from chimpanzee IFNAR1. This human specific trait could contribute to specific immune response in the most optimized manner for humans. Further molecular biological studies on the IFNAR1 will help us to gain insights into the molecular implication of species-specific host-pathogen interaction in primate evolution.

• Journal of Microbiology and Biotechnology
• /
• 제25권8호
• /
• pp.1246-1256
• /
• 2015

Chlamydophila psittaci is an important intracellular pathogen. Persistent infection is an important state of the host-parasite interaction in this chlamydial infection, which plays a significant role in spreading the organism within animal populations and in causing chronic chlamydiosis and serious sequelae. In this study, a C. psittaci persistent infection cell model was induced by penicillin G, and real-time quantitative PCR was used to study the transcriptional levels of 10 C. psittaci genes (dnaA, dnaK, ftsW, ftsY, grpE, rpsD, incC, omcB, CPSIT_0846, and CPSIT_0042) in acute and penicillin-G-induced persistent infection cultures. Compared with the acute cultures, the penicillin-G-treated cultures showed a reduced chlamydial inclusion size and a significantly decreased number of elementary body particles. Additionally, some enlarged aberrant reticulate body particles were present in the penicillin-G-treated cultures but not the acute ones. The expression levels of genes encoding products for cell division (FtsW, FtsY) and outer membrane protein E encoding gene (CPSIT_0042) were downregulated (p < 0.05) from 6 h post-infection onward in the persistent infection cultures. Also from 6 h post-infection, the expression levels of DnaA, DnaK, IncC, RpsD, GrpE, and CPSIT_0846 were upregulated (p < 0.05); however, the expression level of OmcB in the persistent infection was< almost the same as that in the acute infection (p > 0.05). These results provide new insight regarding molecular activities that accompany persistence of C. psittaci, which may play important roles in the pathogenesis of C. psittaci infection.

• Journal of Microbiology and Biotechnology
• /
• 제21권7호
• /
• pp.679-685
• /
• 2011

Xanthomonas oryzae pv. oryzae (Xoo) produces a putative effector, XoAvrBs2. We expressed XoAvrBs2 homologously in Xoo with a TAP-tag at the C-terminus to enable quantitative analysis of protein expression and secretion. Addition of rice leaf extracts from both Xoo-sensitive and Xoo-resistant rice cultivars to the Xoo cells induced expression of the XoAvrBs2 gene at the transcriptional and translational levels, and also stimulated a remarkable amount of XoAvrBs2 secretion into the medium. In a T3SS-defective Xoo mutant strain, secretion of the TAPtagged XoAvrBs2 was blocked. Thus, we elucidated the transcriptional and translational expressions of the XoAvrBs2 gene in Xoo was induced in vitro by the interaction with rice and the induced secretion of XoAvrBs2 was T3SSdependent. It is the first report to measure the homologous expression and secretion of XoAvrBs2 in vitro by rice leaf extract. Our system for the quantitative analysis of effector protein expression and secretion could be generally used for the study of host-pathogen interactions.

• The Plant Pathology Journal
• /
• 제38권1호
• /
• pp.1-11
• /
• 2022

Fusarium culmorum is one of the most important causal agents of root rot of wheat. In this study, 10 F. culmorum isolates were collected from farms located in five agro-ecological regions of Morocco. These were used to challenge 20 durum wheat genotypes via artificial inoculation of plant roots under controlled conditions. The isolate virulence was determined by three traits (roots browning index, stem browning index, and severity of root rot). An alpha-lattice design with three replicates was used, and the resulting ANOVA revealed a significant (P < 0.01) effect of isolate (I), genotype (G), and G × I interaction. A total of four response types were observed (R, MR, MS, and S) revealing that different genes in both the pathogen and the host were activated in 53% of interactions. Most genotypes were susceptible to eight or more isolates, while the Moroccan cultivar Marouan was reported resistant to three isolates and moderately resistant to three others. Similarly, the Australian breeding line SSD1479-117 was reported resistant to two isolates and moderately resistant to four others. The ICARDA elites Icaverve, Berghisyr, Berghisyr2, Amina, and Icaverve2 were identified as moderately resistant. Principal component analysis based on the genotypes responses defined two major clusters and two sub-clusters for the 10 F. culmorum isolates. Isolate Fc9 collected in Khemis Zemamra was the most virulent while isolate Fc3 collected in Haj-Kaddour was the least virulent. This work provides initial results for the discovery of differential reactions between the durum lines and isolates and the identification of novel sources of resistance.

• 대한약침학회지
• /
• 제27권2호
• /
• pp.91-100
• /
• 2024

Objectives: Candida albicans is an opportunistic pathogen that occurs as harmless commensals in the intestine, urogenital tract, and skin. It has been influenced by a variety of host conditions and has now evolved as a resistant strain. The aim of this study was thus detect the fluconazole resistant C. albicans from the root caries specimens and to computationally evaluate the interactions of an opaque-phase ABC transporter protein with the Psidium guajava bio-active compounds. Methods: 20 carious scrapings were collected from patients with root caries and processed for the isolation of C. albicans and was screened for fluconazole resistance. Genomic DNA was extracted and molecular characterization of Cdrp1 and Cdrp2 was done by PCR amplification. P. guajava methanolic extract was checked for the antifungal efficacy against the resistant strain of C. albicans. Further in-silico docking involves retrieval of ABC transporter protein and ligand optimization, molinspiration assessment on drug likeness, docking simulations and visualizations. Results: 65% of the samples showed the presence of C.albicans and 2 strains were fluconazole resistant. Crude methanolic extract of P. guajava was found to be promising against the fluconazole resistant strains of C. albicans. In-silico docking analysis showed that Myricetin was a promising candidate with a high docking score and other drug ligand interaction scores. Conclusion: The current study emphasizes that bioactive compounds from Psidium guajava to be a promising candidate for treating candidiasis in fluconazole resistant strains of C. albicans However, further in-vivo studies have to be implemented for the experimental validation of the same in improving the oral health and hygiene.

• 한국미생물·생명공학회지
• /
• 제46권1호
• /
• pp.77-84
• /
• 2018

$CO_2$의 상승은 식물 병원성 미생물의 발병력과 기주 식물의 저항성에 영향을 미칠 것이며, 기주와 병원체의 상호 작용에도 영향을 미칠 것으로 예상된다. 본 연구는 $CO_2$ 상승 환경에서 기주와 병원체간의 상호 작용을 연구하기 위하여 고추(Capsicum annuum)와 세균점무늬병을 유발하는 X. euvesicatoria를 이용하였다. 고추 식물체의 병저항성 관련 유전자인 CaLRR1, CaWRKY1, CaPIK1 그리고 CaPR10 유전자를 quantitative RT-PCR로 분석한 결과 800 ppm에서 CaLRR1, CaPIK1 그리고 PR10 유전자의 발현이 감소하였으며, negative regulator인 CaWRKY1 유전자는 발현이 증가하였다. 400 ppm과 800 ppm의 $CO_2$ 농도에서 이병엽률과 발병도를 확인 한 결과 800 ppm에서 발병도가 증가된 것을 확인하였다. 이들 결과는 미래의 $CO_2$ 농도가 증가 된 환경에서 고추와 고추의 주요 피해 병원균인 X. euvesicatoria에 의한 고추 세균점무늬병의 발병 양상을 이해하는 기초 자료로 활용할 수 있을 것이다.

• 한국응용곤충학회지
• /
• 제24권2호
• /
• pp.65-70
• /
• 1985

수도(水稻) 병해충(病害蟲)의 동시발생(同時發生) 피해(被害)에 관한 연구(硏究)를 위한 기초시험(基礎試驗)으로 벼멸구와 흰등멸구의 복합발생(複合發生)에 따른 서식처선호성(棲息處選好性)을 확인(確認)코저 감수성(感受性) 품종(品種)인 IR22와 저항성(抵抗性)인 IR36(분벽최성기(分蘗最盛期)부터 흰등멸구에도 저항성(抵抗性)임)을 공시(供試)하여 조사(調査)한 결과(結果) 벼품종별(品種別), 생육시기별(生育時期別) 및 발생밀도(發生密度)에 따라 다소(多少) 차이(差異)는 있으나 일반적(一般的)으로 흰등멸구는 대부분(大部分) 벼 윗부분(部分)에 서식(棲息)하였고 벼멸구는 아래부분(部分)에 서식(棲息)함을 알 수 있었다. 두 종(種)의 복합발생시(複合發生時)의 서식선호성(棲息選好性)은 상대종(相對種)의 발생(發生)에 영향(影響)을 받지 않았고 각각 일정(一定)한 서식처(棲息處)를 선호(選好)하는 경향(傾向)이었다. 벼멸구의 생태형(生態型) 2에 대(對)하여 반응(反應)이 다른 다섯가지 품종(品種)(IR22 및 TN1: 감수성(感受性), Triveni 및 ASD7: 중정도저항성(中程度抵抗性), IR42: 抵抗性)을 공시(供試)하여 문고병(紋枯病)과 벼멸구의 동시발생(同時發生)에 의한 피해(被害)를 조사(調査)한 결과(結果) 병해충(病害蟲)의 동시발생(同時發生) 피해(被害)는 병원균(病原菌)이 단독발생(單獨發生)했을 때 보다 문고병(紋枯病)의 발병(發病)을 현저(顯著)히 조장(助長)시켰으며 병증(病症)의 발현속도(發現速度)도 빨랐고 균계생장(菌系生長)도 왕성(旺盛)하여 감염기(感染器)(infection structure)의 형성(形成)도 풍부(豊富)하였다. 벼멸구의 생태형(生態型) 2에 대(對)한 품종(品種) 반응(反應)과는 상관없이 벼멸구와 문고병(紋枯病)과의 동시발생(同時發生)은 벼멸구 단독발생(單獨發生)에 비(比)하여 더 심한 고사현상(枯死現象)(hopperburn)을 일으켰다. 즉 문고병(紋枯病) 병원균(病原菌)과 벼멸구의 동시발생(同時發生)은 상승적(相乘的) 피해(被害)가 나타남을 확인(確認)하였다.

• 식물병연구
• /
• 제23권3호
• /
• pp.249-255
• /
• 2017

벼 도열병은 벼를 재배하는 지역에서는 가장 중요한 병 중 하나이다. 특히, 벼 도열병균은 기주인 벼와 Gene-for-Gene 상호작용이 적용 가능한 대표적인 모델 식물병원성 곰팡이다. 우리나라는 1980년 이래로 벼 도열병균의 레이스를 분석하기 위해 8개의 판별 품종을 이용한 시스템을 구축하여 분류하였다. 그러나 이 판별 품종이 어떤 저항성 유전자를 가지고 있는지에 관해 명확한 정보가 없어 새로운 레이스의 출현이나 병 저항성 붕괴 등에 대하여 과학적인 분석이 어려웠다. 최근 병원균의 레이스와 벼의 저항성 유전자의 상호작용 이해를 돕기 위해 LTH 품종에 단인자 저항성 계통을 각각 다르게 도입한 판별시스템이 개발되었다. 본 연구에서는 우리나라의 1995년부터 2015년까지 분리된 4개의 다른 레이스 KI101, KI201, KI401 및 KJ101로부터 총 50개 균주를 선발하여 LTH 품종에 기반한 단인자 저항성 계통에 접종하여 그 결과를 이전 레이스와 비교 분석해 보았다. 그 결과 한국형 판별시스템으로 분류된 동일 레이스내의 균주들이 단인자 계통에서 서로 다른 반응을 보였다. 이 결과 동일 레이스에 속하는 균주들이 서로 다른 비병원성 유전자를 지닌 것을 의미하며, 더 나아가 새로운 저항성 벼 품종 육종에 유용한 정보를 제공하기 어려울 것으로 추정되었다. 이 연구 결과 현재의 판별시스템과 더불어 단인자 저항성 품종을 통한 판별시스템 도입이 요구되었다. 이 연구 결과는 향후 한국의 판별시스템 개발에 기초 자료로 활용 될 수 있을 것이다.