Archangium gephyra 의 tubulysin 생합성 유전자 분석 = LAnalysis of Tubulysin Biosynthetic Genes in Archangium gephyra

국문 초록 (Abstract)

Tubulysin은 다양한 암세포주에 대해 강한 항암활성을 보이는 점액세균 유래 이차대사 생리활성물질이다. 본 연구에서는 tubulysin을 생산하는 두 균주의 점액세균 Archangium gephyra MEHO_002와 MEHO_004의 유전체 분석을 통해 tubulysin 생합성 유전자들로 추정되는 유전자군을 발견하였으며, 플라스미드 삽입에 의한 유전자 불활성화를 통해 이들 유전자들이 tubulysin 생산과 직접 연관되어 있음을 확인하였다. A. gephyra MEHO_002와 MEHO_004 균주의 tubulysin 생합성 유전자군(tubA~tubF)은 DNA 염기서열이 서로 97% 동일하였으며, 암호화하는 단백질들의 아미노산 서열도 서로 97-100% 유사하였다. MEHO_002와 MEHO_004 균주의 tubulysin 생합성 유전자군은 tubulysin 생산 점액세균으로 알려진 Cystobacter sp. SBCb004의 tubulysin 생합성 유전자군과 DNA 염기서열이 86% 동일하였다. 유전자군의 구성은 tubZ 유전자가 존재하지 않는다는 점을 제외하고는 SBCb004의 tubulysin 생합성 유전자군 구성과 동일하였다. 각 유전자가 암호화하는 단백질의 아미노산 서열은 Cystobacter sp. SBCb004의 tubulysin 생합성 유전자가 암호화하는 단백질들과 88-97% 유사하였으며, 각 단백질들의 도메인 구성도 동일하였다.

다국어 초록 (Multilingual Abstract)

Tubulysins are a group of bioactive secondary metabolites from myxobacteria exhibiting strong anticancer activity against various cancer cell lines. In this study, we describe the identification of putative tubulysin biosynthetic gene clusters (tubA~tubF) in the genome sequences of two tubulysin-producing myxobacterial strains, Archangium gephyra MEHO_002 and MEHO_004. The inactivation of the putative tubulysin biosynthetic genes resulted in a tubulysin-production defect. The DNA sequences of the A. gephyra MEHO_002 and MEHO_004 tubulysin biosynthetic genes were 97% identical, and the amino acid sequences of the encoded proteins shared a similarity of 97-100%. The nucleotide sequences of the tubulysin biosynthetic gene clusters in MEHO_002 and MEHO_004 were 86% identical to that in Cystobacter sp. SBCb004 known as a tubulysin-producing myxobacterium, and the organization of the clusters was identical except for the lack of a tubZ gene in the clusters in MEHO_002 and MEHO_004. The amino acid sequences of the proteins encoded by each gene were 88-97% similar to those encoded by SBCb004, and the domain compositions of the proteins were also identical.

참고문헌 (Reference)

1 이봉수, "분산 돌연변이 점액세균의 분리" 한국미생물·생명공학회 31 (31): 342-347, 2003

2 Blin K, "antiSMASH 5.0: updates to the secondary metabolite genome mining pipeline" 47 : W81-W87, 2019

3 Sasse F, "Tubulysins, new cytostatic peptides from myxobacteria acting on microtubuli. Production, isolation, physico-chemical and biological properties" 53 : 879-885, 2000

4 Courter JR, "Structure-activity relationships of tubulysin analogues" 30 : 127241-, 2020

5 현혜숙, "Screening of Myxobacteria Carrying Tubulysin Biosynthetic Genes" 한국미생물·생명공학회 49 (49): 32-38, 2021

6 Ullrich A, "Pretubulysin, a potent and chemically accessible tubulysin precursor from Angiococcus disciformis" 48 : 4422-4425, 2009

7 Reddy JA, "Pre-clinical evaluation of EC1456, a folatetubulysin anti-cancer therapeutic" 8 : 8943-, 2018

8 Szigetvari NM, "Phase I/II clinical trial of the targeted chemotherapeutic drug, folate-tubulysin, in dogs with naturallyoccurring invasive urothelial carcinoma" 9 : 37042-37053, 2018

9 Johnson M, "NCBI BLAST : a better web interface" 36 : W5-W9, 2008

10 Dömling A, "Myxobacterial epothilones and tubulysins as promising anticancer agents" 9 : 141-147, 2005

11 Sambrook J, "Molecular Cloning: a Laboratory Manual" Cold Spring Harbor Laboratory Press 1989

12 Risinger AL, "Microtubule dynamics as a target in oncology" 35 : 255-261, 2009

13 Khalil MW, "Mechanism of action of tubulysin, an antimitotic peptide from myxobacteria" 7 : 678-683, 2006

14 Steinmetz H, "Isolation, crystal and solution structure determination, and biosynthesis of tubulysins - powerful inhibitors of tubulin polymerization from myxobacteria" 43 : 4888-4892, 2004

15 Sandmann A, "Identification and analysis of the core biosynthetic machinery of tubulysin, a potent cytotoxin with potential anticancer activity" 11 : 1071-1079, 2004

16 Chai Y, "Heterologous expression and genetic engineering of the tubulysin biosynthetic gene cluster using Red/ET recombineering and inactivation mutagenesis" 19 : 361-371, 2012

17 Etienne-Manneville S, "From signaling pathways to microtubule dynamics : the key players" 22 : 104-111, 2010

18 Chai Y, "Discovery of 23 natural tubulysins from Angiococcus disciformis An d48 and Cystobacter SBCb004" 17 : 296-309, 2010

19 Shimkets LJ, "Correlation of energy-dependent cell cohesion with social motility in Myxococcus xanthus" 166 : 837-841, 1986

20 신혜진, "Corallococcus와 Myxococcus 속 점액세균 균주들에 의한 항균 물질의 생산" 한국미생물·생명공학회 41 (41): 44-51, 2013

21 Murray BC, "Chemistry and biology of tubulysins : antimitotic tetrapeptides with activity against drug resistant cancers" 32 : 654-662, 2015

22 Marchler-Bauer A, "CDD : NCBI's conserved domain database" 43 : D222-D226, 2015

23 Kaur G, "Biological evaluation of tubulysin A : a potential anticancer and antiangiogenic natural product" 396 : 235-242, 2006

24 Sasse F, "Argyrins, immunosuppressive cyclic peptides from myxobacteria. I. Production, isolation, physicochemical and biological properties" 55 : 543-551, 2002

25 Selva E, "Antibiotics A21459 A and B, new inhibitors of bacterial protein synthesis. I. Taxonomy, isolation and characterization" 49 : 145-149, 1996