Journal Article
- The cytoplasmic loops of AgrC contribute to the quorum-sensing activity of Staphylococcus aureus
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Qian Huang , Yihui Xie , Ziyu Yang , Danhong Cheng , Lei He , Hua Wang , Qian Liu , Min Li
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J. Microbiol. 2021;59(1):92-100. Published online November 17, 2020
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DOI: https://doi.org/10.1007/s12275-021-0274-x
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Abstract
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In Staphylococcus aureus, the accessory gene regulator (agr)
quorum-sensing system is thought to play an important role
in biofilm formation. The histidine kinase AgrC is one of the
agr system components and activated by the self-generated
auto-inducing peptide (AIP), which is released continuously
into the extracellular environment during bacterial growth.
The extracellular loops (Extra-loops) of AgrC are crucial for
AIP binding. Here, we reported that the cytoplasmic loops
(Cyto-loops) of AgrC are also involved in Agr activity. We
identified S. aureus ST398 clinical isolates containing a naturally
occurring single amino acid substitution (lysine to isoleucine)
at position 73 of an AgrC Cyto-loop that exhibited
significantly stronger biofilm formation and decreased Agr
activity compared to the wild-type strain. A constructed strain
containing the K73I point mutation in AgrC Cyto-loop continued
to show a growth dependent induction of the agr system,
although the growth dependent induction was delayed
by about 6 h compared to the wild-type. In addition, a series
of strains containing deletion mutants of the AgrC Cyto- and
Extra-loops were constructed and revealed that the removal
of the two Cyto-loops and Extra-loops 2 and 3 totally abolished
the Agr activity and the growth-dependence on the agr
system induction. Remarkably, the Extra-loop 1 deletion did
not affect the Agr activity. In conclusion, the AgrC Cyto-loops
play a crucial role in the S. aureus quorum-sensing activity.
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- Emerging trends in the inhibition of bacterial molecular communication: An overview
B.R. Maha Swetha, M. Saravanan, Prakash Piruthivraj
Microbial Pathogenesis.2024; 186: 106495. CrossRef - In vitro and in silico evaluation of anti-quorum sensing activity of marine red seaweeds-Portieria hornemannii and Halymenia dilatata
Prakash Piruthivraj, B.R. Maha Swetha, A. Anita Margret, A. Sherlin Rosita, Parthasarathi Rengasamy, Rajapandiyan Krishnamoorthy, Mansour K. Gatasheh, Khalid Elfaki Ibrahim, Sekhu Ansari, Natesan Vijayakumar
Journal of King Saud University - Science.2024; 36(6): 103188. CrossRef - Commensal Staphylococcus epidermidis Defends against Staphylococcus aureus through SaeRS Two-Component System
Nadira Nurxat, Lili Wang, Qichen Wang, Shujing Li, Chen Jin, Yaran Shi, Ayjiamali Wulamu, Na Zhao, Yanan Wang, Hua Wang, Min Li, Qian Liu
ACS Omega.2023; 8(20): 17712. CrossRef - Biofilm and Small Colony Variants—An Update on Staphylococcus aureus Strategies toward Drug Resistance
Henan Guo, Yucui Tong, Junhao Cheng, Zaheer Abbas, Zhongxuan Li, Junyong Wang, Yichen Zhou, Dayong Si, Rijun Zhang
International Journal of Molecular Sciences.2022; 23(3): 1241. CrossRef
Retracted Publication
- Cryptic prophages in a blaNDM-1-bearing plasmid increase bacterial survival against high NaCl concentration, high and low temperatures, and oxidative and immunological stressors
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So Yeon Kim , Kwan Soo Ko
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J. Microbiol. 2020;58(6):483-488. Published online March 28, 2020
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DOI: https://doi.org/10.1007/s12275-020-9605-6
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50
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Abstract
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In this study, we investigated the effect of cryptic prophage
regions in a blaNDM-1-bearing plasmid, which was identified in
a patient from South Korea, on the survival of bacteria against
adverse environmental conditions. First, we conjugated the
intact plasmid and plasmids with deleted cryptic prophages
into Escherichia coli DH5α. The E. coli transconjugants carrying
the plasmid with intact cryptic prophages showed increased
survival during treatment with a high concentration
of NaCl, high and low temperatures, an oxidative stressor
(H2O2), and an immunological stressor (human serum). By
contrast, the transconjugants carrying the plasmid with a
single-cryptic prophage knockout did not show any change
in survival rates. mRNA expression analyses revealed that the
genes encoding sigma factor proteins were highly upregulated
by the tested stressors and affected the expression of
various proteins (antioxidant, cell osmosis-related, heat shock,
cold shock, and universal stress proteins) associated with the
specific defense against each stress. These findings indicate
that a bacterial strain carrying a plasmid with intact carbapenemase
gene and cryptic prophage regions exhibited an increased
resistance against simulated environmental stresses,
and cryptic prophages in the plasmid might contribute to this
enhanced stress resistance. Our study indicated that the coselection
of antibiotic resistance and resistance to other stresses
may help bacteria to increase survival rates against adverse
environments and disseminate.
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- Uncovering the virome and its interaction with antibiotic resistome during compost fertilization
Qingxia Zhang, Lei Zhou, Yilong Zhao, Shuhong Gao, Yanjun Yang, Qingyun Chen, Wenhui Li, Qi Qi, Qiang Dong, Jiesi Lei, Xue Guo, Qun Gao, Yunfeng Yang
Journal of Hazardous Materials.2023; 457: 131763. CrossRef - Regulator of RNase E activity modulates the pathogenicity of Salmonella Typhimurium
Jaejin Lee, Eunkyoung Shin, Ji-Hyun Yeom, Jaeyoung Park, Sunwoo Kim, Minho Lee, Kangseok Lee
Microbial Pathogenesis.2022; 165: 105460. CrossRef - Presence and Persistence of Putative Lytic and Temperate Bacteriophages in Vaginal Metagenomes from South African Adolescents
Anna-Ursula Happel, Christina Balle, Brandon S. Maust, Iyaloo N. Konstantinus, Katherine Gill, Linda-Gail Bekker, Rémy Froissart, Jo-Ann Passmore, Ulas Karaoz, Arvind Varsani, Heather Jaspan
Viruses.2021; 13(12): 2341. CrossRef - Regulator of ribonuclease activity modulates the pathogenicity of Vibrio vulnificus
Jaejin Lee, Eunkyoung Shin, Jaeyeong Park, Minho Lee, Kangseok Lee
Journal of Microbiology.2021; 59(12): 1133. CrossRef
Journal Articles
- A histone deacetylase, MoHOS2 regulates asexual development and virulence in the rice blast fungus
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Jongjune Lee , Jae-Joon Lee , Junhyun Jeon
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J. Microbiol. 2019;57(12):1115-1125. Published online November 22, 2019
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DOI: https://doi.org/10.1007/s12275-019-9363-5
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48
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15
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16
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Abstract
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Histone acetylation/deacetylation represent a general and
efficient epigenetic mechanism through which fungal cells control
gene expression. Here we report developmental requirement
of MoHOS2-mediated histone deacetylation (HDAC)
for the rice blast fungus, Magnaporthe oryzae. Structural similarity
and nuclear localization indicated that MoHOS2 is an
ortholog of Saccharomyces cerevisiae Hos2, which is a member
of class I histone deacetylases and subunit of Set3 complex.
Deletion of MoHOS2 led to 25% reduction in HDAC
activity, compared to the wild-type, confirming that it is a
bona-fide HDAC. Lack of MoHOS2 caused decrease in radial
growth and impinged dramatically on asexual sporulation.
Such reduction in HDAC activity and phenotypic defects of
ΔMohos2 were recapitulated by a single amino acid change
in conserved motif that is known to be important for HDAC
activity. Expression analysis revealed up-regulation of MoHOS2
and concomitant down-regulation of some of the key genes
involved in asexual reproduction under sporulation-promoting
condition. In addition, the deletion mutant exhibited defect
in appressorium formation from both germ tube tip and
hyphae. As a result, ΔMohos2 was not able to cause disease
symptoms. Wound-inoculation showed that the mutant is
compromised in its ability to grow inside host plants as well.
We found that some of ROS detoxifying genes and known
effector genes are de-regulated in the mutant. Taken together,
our data suggest that MoHOS2-dependent histone deacetylation
is pivotal for proper timing and induction of transcription
of the genes that coordinate developmental changes
and host infection in M. oryzae.
-
Citations
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- Glsirt1-mediated deacetylation of GlCAT regulates intracellular ROS levels, affecting ganoderic acid biosynthesis in Ganoderma lucidum
Jing Han, Lingshuai Wang, Xin Tang, Rui Liu, Liang Shi, Jing Zhu, Mingwen Zhao
Free Radical Biology and Medicine.2024; 216: 1. CrossRef - Histone (de)acetylation in epigenetic regulation of Phytophthora pathobiology
Yufeng Guan, Joanna Gajewska, Jolanta Floryszak‐Wieczorek, Umesh Kumar Tanwar, Ewa Sobieszczuk‐Nowicka, Magdalena Arasimowicz‐Jelonek
Molecular Plant Pathology.2024;[Epub] CrossRef - FolSas2 is a regulator of early effector gene expression during Fusarium oxysporum infection
Limin Song, Yalei Wang, Fahui Qiu, Xiaoxia Li, Jingtao Li, Wenxing Liang
New Phytologist.2024;[Epub] CrossRef - Regulatory roles of epigenetic modifications in plant-phytopathogen interactions
Zeng Tao, Fei Yan, Matthias Hahn, Zhonghua Ma
Crop Health.2023;[Epub] CrossRef - The additional PRC2 subunit and Sin3 histone deacetylase complex are required for the normal distribution of H3K27me3 occupancy and transcriptional silencing in Magnaporthe oryzae
Chuyu Lin, Zhongling Wu, Huanbin Shi, Jinwei Yu, Mengting Xu, Fucheng Lin, Yanjun Kou, Zeng Tao
New Phytologist.2022; 236(2): 576. CrossRef - Regulatory Roles of Histone Modifications in Filamentous Fungal Pathogens
Yiling Lai, Lili Wang, Weilu Zheng, Sibao Wang
Journal of Fungi.2022; 8(6): 565. CrossRef - Polycomb Repressive Complex 2-Mediated H3K27 Trimethylation Is Required for Pathogenicity in Magnaporthe oryzae
Zhongling Wu, Jiehua Qiu, Huanbin Shi, Chuyu Lin, Jiangnan Yue, Zhiquan Liu, Wei Xie, Naweed I. Naqvi, Yanjun Kou, Zeng Tao
Rice Science.2022; 29(4): 363. CrossRef - Protein acetylation and deacetylation in plant‐pathogen interactions
Jing Wang, Chao Liu, Yun Chen, Youfu Zhao, Zhonghua Ma
Environmental Microbiology.2021; 23(9): 4841. CrossRef - Emerging Roles of Posttranslational Modifications in Plant-Pathogenic Fungi and Bacteria
Wende Liu, Lindsay Triplett, Xiao-Lin Chen
Annual Review of Phytopathology.2021; 59(1): 99. CrossRef - Fungal Lysine Deacetylases in Virulence, Resistance, and Production of Small Bioactive Compounds
Ingo Bauer, Stefan Graessle
Genes.2021; 12(10): 1470. CrossRef - A Histone Deacetylase, Magnaporthe oryzae RPD3, Regulates Reproduction and Pathogenic Development in the Rice Blast Fungus
Song Hee Lee, Mohamed El-Agamy Farh, Jaejoon Lee, Young Taek Oh, Eunbyeol Cho, Jiyeun Park, Hokyoung Son, Junhyun Jeon, Antonio Di Pietro
mBio.2021;[Epub] CrossRef - The Histone Deacetylases MoRpd3 and MoHst4 Regulate Growth, Conidiation, and Pathogenicity in the Rice Blast Fungus Magnaporthe oryzae
Chaoxiang Lin, Xue Cao, Ziwei Qu, Shulin Zhang, Naweed I. Naqvi, Yi Zhen Deng, Aaron P. Mitchell
mSphere.2021;[Epub] CrossRef - Histone Acetyltransferases and Deacetylases Are Required for Virulence, Conidiation, DNA Damage Repair, and Multiple Stresses Resistance of Alternaria alternata
Haijie Ma, Lei Li, Yunpeng Gai, Xiaoyan Zhang, Yanan Chen, Xiaokang Zhuo, Yingzi Cao, Chen Jiao, Fred G. Gmitter, Hongye Li
Frontiers in Microbiology.2021;[Epub] CrossRef - Function of PoLAE2, a laeA homolog, in appressorium formation and cAMP signal transduction in Pyricularia oryzae
Pradabrat Prajanket, Kim-Chi Thi Vu, Jun Arai, Worawan Sornkom, Ayumi Abe, Teruo Sone
Bioscience, Biotechnology, and Biochemistry.2020; 84(11): 2401. CrossRef - A Histone Deacetylase, MoHDA1 Regulates Asexual Development and Virulence in the Rice Blast Fungus
Taehyun Kim, Song Hee Lee, Young Taek Oh, Junhyun Jeon
The Plant Pathology Journal.2020; 36(4): 314. CrossRef - Protein Acetylation/Deacetylation: A Potential Strategy for Fungal Infection Control
Junzhu Chen, Qiong Liu, Lingbing Zeng, Xiaotian Huang
Frontiers in Microbiology.2020;[Epub] CrossRef
- Construction of a genetically modified T7Select phage system to express the antimicrobial peptide 1018
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David J. Lemon , Matthew K. Kay , James K. Titus , April A. Ford , Wen Chen , LCDR Nicholas J. Hamlin , Yoon Y. Hwang
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J. Microbiol. 2019;57(6):532-538. Published online May 27, 2019
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DOI: https://doi.org/10.1007/s12275-019-8686-6
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46
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23
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23
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Abstract
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Bacteriophage therapy was an ascendant technology for combating
bacterial infections before the golden age of antibiotics,
but the therapeutic potential of phages was largely ignored
after the discovery of penicillin. Recently, with antibioticresistant
infections on the rise, these phages are receiving renewed
attention to combat problematic bacterial infections.
Our approach is to enhance bacteriophages with antimicrobial
peptides, short peptides with broad-spectrum antibiotic or
antibiofilm effects. We inserted coding sequences for 1018,
an antimicrobial peptide previously shown to be an effective
broad-spectrum antimicrobial and antibiofilm agent, or the
fluorescent marker mCherry, into the T7Select phage genome.
Transcription and production of 1018 or mCherry began
rapidly after E. coli cultures were infected with genetically modified
phages. mCherry fluorescence, which requires a 90 min
initial maturation period, was observed in infected cultures
after 2 h of infection. Finally, we tested phages expressing 1018
(1018 T7) against bacterial planktonic cultures and biofilms,
and found the 1018 T7 phage was more effective than the
unmodified T7Select phage at both killing planktonic cells and
eradicating established biofilms, validating our phage-driven
antimicrobial peptide expression system. The combination
of narrow-spectrum phages delivering relatively high local
doses of broad-spectrum antimicrobials could be a powerful
method
to combat resistant infections. The experiments we
describe prove this combination is feasible in vitro, but further
testing and optimization are required before genetically modified
phages are ready for use in vivo.
-
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Susanne Meile, Jiemin Du, Matthew Dunne, Samuel Kilcher, Martin J Loessner
Current Opinion in Virology.2022; 52: 182. CrossRef - Gold nanoparticle-DNA aptamer-assisted delivery of antimicrobial peptide effectively inhibits Acinetobacter baumannii infection in mice
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Journal of Microbiology.2022; 60(1): 128. CrossRef - How Good are Bacteriophages as an Alternative Therapy to Mitigate Biofilms of Nosocomial Infections
Aditi Singh, Sudhakar Padmesh, Manish Dwivedi, Irena Kostova
Infection and Drug Resistance.2022; Volume 15: 503. CrossRef - Comparative Analysis of NanoLuc Luciferase and Alkaline Phosphatase Luminescence Reporter Systems for Phage-Based Detection of Bacteria
Shalini Wijeratne, Arindam Bakshi, Joey Talbert
Bioengineering.2022; 9(9): 479. CrossRef - Construction and Characterization of T7 Bacteriophages Harboring Apidaecin-Derived Sequences
Tobias Ludwig, Ralf Hoffmann, Andor Krizsan
Current Issues in Molecular Biology.2022; 44(6): 2554. CrossRef - Genetic and Chemical Engineering of Phages for Controlling Multidrug-Resistant Bacteria
Dingming Guo, Jingchao Chen, Xueyang Zhao, Yanan Luo, Menglu Jin, Fenxia Fan, Chaiwoo Park, Xiaoman Yang, Chuqing Sun, Jin Yan, Weihua Chen, Zhi Liu
Antibiotics.2021; 10(2): 202. CrossRef - Antibiofilm activity of host defence peptides: complexity provides opportunities
Robert E. W. Hancock, Morgan A. Alford, Evan F. Haney
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Stephen Amankwah, Kedir Abdusemed, Tesfaye Kassa
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Advances in Microbiology.2021; 10(01): 30. CrossRef - Phages for Biofilm Removal
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Reviews
- MINIREVIEW] Multilayered regulations of RIG-I in the anti-viral signaling pathway
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Nari Kim , Hesung Now , Nhung T.H. Nguyen , Joo-Yeon Yoo
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J. Microbiol. 2016;54(9):583-587. Published online August 31, 2016
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DOI: https://doi.org/10.1007/s12275-016-6322-2
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46
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Abstract
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RIG-I is a cytosolic receptor recognizing virus-specific RNA
structures and initiates an antiviral signaling that induces the
production of interferons and proinflammatory cytokines.
Because inappropriate RIG-I signaling affects either viral
clearance or immune toxicity, multiple regulations of RIG-I
have been investigated since its discovery as the viral RNA
detector. In this review, we describe the recent progress in
research on the regulation of RIG-I activity or abundance.
Specifically, we focus on the mechanism that modulates RIGI-
dependent antiviral response through post-translational
modifications of or protein-protein interactions with RIG-I.
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Ni Zhong, Chen Wang, Guangxiu Weng, Ting Ling, Liangguo Xu
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Xueren Ouyang, Yuning Zeng, Xiaotao Jiang, Hua Xu, Yile Ning, Ming Yi
BioMed Research International.2021; 2021: 1. CrossRef - HSPBP1 facilitates cellular RLR-mediated antiviral response by inhibiting the K48-linked ubiquitination of RIG-I
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- REVIEW] All about that fat: Lipid modification of proteins in Cryptococcus neoformans
-
Felipe H. Santiago-Tirado , Tamara L. Doering
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J. Microbiol. 2016;54(3):212-222. Published online February 27, 2016
-
DOI: https://doi.org/10.1007/s12275-016-5626-6
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55
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Abstract
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Lipid modification of proteins is a widespread, essential process
whereby fatty acids, cholesterol, isoprenoids, phospholipids,
or glycosylphospholipids are attached to polypeptides.
These hydrophobic groups may affect protein structure, function,
localization, and/or stability; as a consequence such modifications
play critical regulatory roles in cellular systems.
Recent advances in chemical biology and proteomics have
allowed the profiling of modified proteins, enabling dissection
of the functional consequences of lipid addition. The
enzymes that mediate lipid modification are specific for both
the lipid and protein substrates, and are conserved from fungi
to humans. In this article we review these enzymes, their substrates,
and the processes involved in eukaryotic lipid modification
of proteins. We further focus on its occurrence in
the fungal pathogen Cryptococcus neoformans, highlighting
unique features that are both relevant for the biology of the
organism and potentially important in the search for new
therapies.
-
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Daniel P Agustinho, Liza C Miller, Lucy X Li, Tamara L Doering
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Research Support, Non-U.S. Gov't
- Functional Characterization of the Genes tauO, tauK, and tauI in the Biosynthesis of Tautomycetin
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Fen Wang , Rixiang Kong , Bo Liu , Jing Zhao , Rongguo Qiu , Li Tang
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J. Microbiol. 2012;50(5):770-776. Published online November 4, 2012
-
DOI: https://doi.org/10.1007/s12275-012-2154-x
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Abstract
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Tautomycetin is a specific protein phosphatase I inhibitor. In an effort to elucidate the biosynthetic mechanism of tautomycetin, we inactivated genes of the tautomycetin biosynthetic gene cluster, tauI, tauO, and tauK, which encode for putative P450 oxidase, citryl-CoA lyase, and esterase enzymes, respectively. The mutant STQ0606 (ΔtauO) did not produce any detectable amount of tautomycetin intermediates but could convert dialkylmaleic anhydride to tautomycetin, strongly indicating that TauO was involved in dialkylmaleic anhydride biosynthesis. STQ1211 (ΔtauK) accumulated dialkylmaleic anhydride, whereas the cofermentation of STQ1211 (ΔtauK) and STQ0606 (ΔtauO) restored the production of tautomycetin. Together, these results suggest that TauK was responsible for the conjugation of dialkylmaleic anhydride and the polyketide moiety in tautomycetin biosynthesis. The disruption of tauI resulted in the accumulation of 5-des-ketotautomycetin, revealing that TauI was responsible for the oxidation at C5 as the last step. Although the shunt pathways were involved in the biosynthesis of tautomycetin, the main post-polyketide synthase tailoring steps were dehydration, decarboxylation and oxidation, taking place consecutively. This study allowed us to predict the biosynthesis of tautomycetin more accurately and provided novel insights into the mechanism of the biosynthesis of tautomycetin.
Review
- Biosynthesis, Modification, and Biodegradation of Bacterial Medium-Chain-Length Polyhydroxyalkanoates
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Do Young Kim , Hyung Woo Kim , Moon Gyu Chung , Young Ha Rhee
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J. Microbiol. 2007;45(2):87-97.
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DOI: https://doi.org/2528 [pii]
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Abstract
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Medium-chain-length polyhydroxyalkanoates (MCL-PHAs), which have constituents with a typical chain length of C6-C14, are polyesters that are synthesized and accumulated in a wide variety of Gram-negative bacteria, mainly pseudomonads. These biopolyesters are promising materials for various applications because they have useful mechanical properties and are biodegradable and biocompatible. The versatile metabolic capacity of some Pseudomonas spp. enables them to synthesize MCL-PHAs that contain various functional substituents; these MCL-PHAs are of great interest because these functional groups can improve the physical properties of the polymers, allowing the creation of tailor-made products. Moreover, some functional substituents can be modified by chemical reactions to obtain more useful groups that can extend the potential applications of MCL-PHAs as environmentally friendly polymers and functional biomaterials for use in biomedical fields. Although MCL-PHAs are water-insoluble, hydrophobic polymers, they can be degraded by microorganisms that produce extracellular MCL-PHA depolymerase. MCL-PHA-degraders are relatively uncommon in natural environments and, to date, only a limited number of MCL-PHA depolymerases have been investigated at the molecular level. All known MCL-PHA depolymerases share a highly significant similarity in amino acid sequences, as well as several enzymatic characteristics. This paper reviews recent advances in our knowledge of MCL-PHAs, with particular emphasis on the findings by our research group.
- Role of the Amino Acid Residued in the Catalysis of Catechol 2,3-dioxygenase from Pseudomonas putida SU10 as Probed by Chemical Modification and Random Mutagenesis
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Park, Sun Jung , park, Jin Mo , Lee, Byeong Jae , Min, Kyung Hee
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J. Microbiol. 1997;35(4):300-308.
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Abstract
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The catechol 2,3-dioxygenase (C23O) encoded by the Pseudomonas putida xylE gene was over-produced in Escherichia coli and purified to homogeneity. The activity of the C23O required the reduced form of the Fe(II) ion since the enzyme was highly susceptible to inactivation with hydrogen perocide but reactivated with the addition of ferrous sulfate in conjunction with ascorbic acid. The C23O activity was abolished by treatment with the chemical reagents, diethyl-pyrocarbonate (DEPC), tetranitromethane (TNM), and 1-cyclohexy1-3-(2-morpholinoethyl) car-bodiimidemetho-ρ-toluenesulfontate (CMC), which are modifying reagents of histidine, tyrosine and glutamic acid, respectively. These results suggest that histidine, tyrosine and glutamic acid residues may be good active sites for the enzyme activity. These amino acid residues are conserved residues may be good active sites for the enzyme activity. These amino acid residues are conserved residues among several extradion dioxygenases and have the chemical potential to serveas ligands for Fe(II) coordination. Analysis of random point mutants in the C23O gene derived by PCR technique revealed that the mutated positions of two mutants, T179S and S211R, were located near the conserved His165 amd Hos217 residues, respectively. This finding indicates that these two positions, along with the conserved histidine residues, are specially effective regions for the enzyme function.
- Chemical midification of Cytosine Deaminase from Aspergillus fumigatus
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Yu, Tae Shick , Kim, Jung , Kim, Hyun Soo
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J. Microbiol. 1998;36(1):39-42.
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Abstract
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Essential amino acids involved in the catalytic mechanisms of cytosine deaminase from Aspergillus fumigatus IFO 5840 were determined by chemical modification studies. The enzyme was perfectly inhibited by N-bromosuccinimide, chloramines-T, pyridoxal-5’-phosphate, and p-chloromercuribenzoate. It was strongly inhibited by phenylmethyl sulfonyl fluoride, and weakly inhibited by phenylglyoxal. The inactivation of the enzyme activity by p-CMB was reversed by sulfhydryl reagents. Furthermore, activities inhibited by chloramines-T, pyridoxal-5’-phosphate, results, we speculate that tryptophan, methionine, lysine and cysteine residues are located in ornear the active center of the cytosine deaminase, while a serine is indirectly involved I the enzyme activity.
- Intracellular Posttranslational Modification of Aspartyl Proteinase of Candida albicans and the Role of the Glycan Region of the Enzyme
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Byoung-Kuk Na , Chul-Yong Song
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J. Microbiol. 2000;38(4):218-223.
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Abstract
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Using two drugs, tunicamycin and brefeldin A, which affect protein processing, we investigated the intracellular processing mechanism of secreted aspartyl proteinase 1 (SAP1) of Candida albicans. Three intracellular forms of SAP1 were detected by immunoblotting using monoclonal antibody (MAb) CAP1. Their molecular weights were approximately 40, 41 and 45 kDa, respectively. The 41 kDa protein is a glycoprotein and may be the same as the extracellular form judging by its molecular mass. The 40 kDa protein was the unglycosylated form and its molecular mass coincided with deglycosylated SAP1 and the 45 kDa protein was also the unglycosylated form. Neither the 40 and 45 kDa proteins were detected in the culture supernatant of C. albicans. These suggested that the 40 and 45 kDa proteins might be intracellular precursor forms of SAP1. These results show that SAP1 is translated as a 45 kDa precusor form in the endoplasmic reticulum and the 45 kDa precursor form undergoes proteolytic cleavage after translocation into the Golgi apparatus, generating the 40 kDa precursor form. This 40 kDa precursor is converted into a 41 kDa mature form through glycosylation in the Golgi apparatus. The mature form of the 41 kDa protein is sorted into secretory vesicles and finally released into the extracellular space through membrane fusion. When the glycan region of SAP1 was digested with N-glycosidase F, both stability and activity of the enzyme decreased. These results indicate that the glycan attached to the enzyme may, at least in part, be related to enzyme stability and activity.
- Characterization of Cell Wall Proteins from the soo1-1/ret1-1 Mutant of Saccharomyces cerevisiae
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Dong-Won Lee , Ki-Hyun Kim , Se-Chul Chun , Hee-Moon Park
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J. Microbiol. 2002;40(3):219-223.
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Abstract
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In order to investigate the function of Soo1p/[alpha]-COP during post-translational modification and intracellular transport of cell wall proteins in Saccharomyces cerevisiae, cell wall proteins from the soo1-1/ret1-1 mutant cells were analyzed. SDS-PAGE analysis of biotin labeled cell wall proteins suggested that the soo1-1 mutation impairs post-translational modification of cell wall proteins, such as N- and/or O-glycosylation. Analysis of cell wall proteins with antibodies against [beta]-1,3-glucan and [beta]-1,6-glucan revealed alteration of the linkage between cell wall proteins and [beta]-glucans in the soo1-1 mutant cells. Compositional sugar analysis of the cell wall proteins also suggested that the soo1-1 mutation impairs glycosylation of cell wall protein in the ER, which is crucial for the maintenance of cell wall integrity.