• 제목/요약/키워드: ribosomal

검색결과 1,082건 처리시간 0.024초

Ribosomal Crystallography: Peptide Bond Formation, Chaperone Assistance and Antibiotics Activity

  • Yonath, Ada
    • Molecules and Cells
    • /
    • 제20권1호
    • /
    • pp.1-16
    • /
    • 2005
  • The peptidyl transferase center (PTC) is located in a protein free environment, thus confirming that the ribosome is a ribozyme. This arched void has dimensions suitable for accommodating the 3'ends of the A-and the P-site tRNAs, and is situated within a universal sizable symmetry-related region that connects all ribosomal functional centers involved in amino-acid polymerization. The linkage between the elaborate PTC architecture and the A-site tRNA position revealed that the A-to P-site passage of the tRNA 3'end is performed by a rotatory motion, which leads to stereochemistry suitable for peptide bond formation and for substrate mediated catalysis, thus suggesting that the PTC evolved by genefusion. Adjacent to the PTC is the entrance of the protein exit tunnel, shown to play active roles in sequence-specific gating of nascent chains and in responding to cellular signals. This tunnel also provides a site that may be exploited for local co-translational folding and seems to assist in nascent chain trafficking into the hydrophobic space formed by the first bacterial chaperone, the trigger factor. Many antibiotics target ribosomes. Although the ribosome is highly conserved, subtle sequence and/or conformational variations enable drug selectivity, thus facilitating clinical usage. Comparisons of high-resolution structures of complexes of antibiotics bound to ribosomes from eubacteria resembling pathogens, to an archaeon that shares properties with eukaryotes and to its mutant that allows antibiotics binding, demonstrated the unambiguous difference between mere binding and therapeutical effectiveness. The observed variability in antibiotics inhibitory modes, accompanied by the elucidation of the structural basis to antibiotics mechanism justifies expectations for structural based improved properties of existing compounds as well as for the development of novel drugs.

Brevibacterium ammoniagenes의 30S 리보좀 단백질 S1을 코드하는 유전자의 염기서열 (Nucleotide Sequence of the Putative Gene Encoding 30S Ribosomal Protein S1 from Brevibacterium ammoniagenes)

  • 윤기홍;이미성;오영필;최정호
    • 한국미생물·생명공학회지
    • /
    • 제28권3호
    • /
    • pp.147-151
    • /
    • 2000
  • Brevibacterium ammoniagenes 염색체상에서 phosphotrans-ferase system의 glucose permease를 코드하는 ptsG 유전자와 인접한 지역의 염기서열을 결정한 결돠 1,467 nucleo-tides로 구성된 1개의 open reading frame(ORF)이 발견되었고 이것은 489 아미노산 잔기로 구성되는 단백질을 코드하는 것으로추정된다. 이러한 ORF로부터 추정된 단백질의 아미노산 잔기배열을 분석한 결과 30S 리보좀을 구성하는 단백질중의 하나인 S1과 상동성이 높은 것으로 나타났는데 특히 Mycobacterium tuberculosis M. leprae와 Srepto-myces coelicola의 S1단백질의 아미노산 잔기배열과 각각 83%, 74%m, 77%의 매우 높은 상동성을 보였으며 Escherichia coli의 것과도 약 40%의 상동성을 보였다 이로보아 B.ammoniagenes 염색체상에서 ptsG 유전자와 인접한 지역에 존재하는 ORF는 리보좀 단백질 S1의 유전자로 추정된다. 또한 이들은 염색체상에서 동일한 방향으로 판독되며 S1의 유전자가 ptsG의 위 지역으로 266 nucleotides 떨어져 존재하고 있다.

  • PDF

Amplified Ribosomal DNA Restriction Analysis (ARDRA) 방법을 이용한 국내 분리 Acinetobacter calcoaceticus - Acinetobacter baumannii Complex 균주의 유전자종 동정 (Genomic Species Identification of Acinetobacter calcoaceticus - Acinetobacter baumannii Complex Strains by Amplified Ribosomal DNA Restriction Analysis (ARDRA))

  • 오재영;조재위;박종천;이제철
    • 대한미생물학회지
    • /
    • 제35권1호
    • /
    • pp.69-76
    • /
    • 2000
  • Members of the genus Acinetobacter are recognized as newer pathogens of the nosocomial infection with an increasing frequency in recent years. Strains that belonged to A. calcoaceticus A. baumannii complex (genomic species 1, 2, 3, and 13TU) were major groups associated with nosocomial infection. Phenotypic identification was unreliable and laborious method to classify Acinetobacter strains into 19 genomic species. Rapid and reliable identification of clinical isolates is essential to diagnosis and epidemiology of Acinetobacter. We investigated the suitability of amplified ribosomal DNA restriction analysis (ARDRA) to identify genomic species of 131 Acinetobacter isolates. The 16S rRNA genes (ribosomal DNA) were enzymatically amplified and the amplified PCR products were restricted independently with the enzymes, AluI, CfoI, and MboI. Genomic species of Acinetobacter was classified by the combinations of restriction patterns. The analysis was showed that restriction profiles were characteristic for each genomic species. One hundred fourteen isolates were identified as A. baumannii, twelve were identified as genomic species 13TU, and one was identified as genomic species 3. Four isolates were found to be unknown organisms. All of the isolates which were identified to A. baumannii by phenotypic tests were completely discriminated into A. baumannii and genomic species 13TU by ARDRA. This study demonstrates that ARDRA is a rapid and simple techniques for the identification of Acinetobacter species according to the genomic species.

  • PDF

진딧물의 전 ribosomal RNA 염기배열 (Nucleotide Sequences of an Aphid ribosomal RNA Unit)

  • 권태영;안승락;송철;박종균;김영섭;황재삼;권오유
    • 생명과학회지
    • /
    • 제8권1호
    • /
    • pp.32-39
    • /
    • 1998
  • 진딧물이 하나의 ribosomal RNA 유전자(rDNA)단위는 총 길이가 13,061bp이며 총 G/C비율은 59%이다. 그것을 구성하고 있는 각 영역의 길이와 G/C비율은 다음과 같다. 5’ETS는 G/C비율이 69%이고 843bp이다 . 18S rRNA 는 2,469bp이며 G/C비율은 59%이다. ITS I길이는 229bp이며 70%의 G/C비율이다. 5.8S rRNA는 160bp이며 63%의 G/C비율이다. ITS II는 325bp이며 70%dml G/C비율이다. 28S rRNA는 4, 147bp이고 60%의 G/C비율이다. IGS는 4,888bp로 55%의 G/C비율이다.

  • PDF

진핵생물과 원핵생물의 미토콘드리아 관련 보존적 유전자 비교 (Comparison of Mitochondria-related Conserved Genes in Eukaryotes and Prokaryotes)

  • 이동근
    • 생명과학회지
    • /
    • 제24권7호
    • /
    • pp.791-797
    • /
    • 2014
  • 원핵과 진핵생물에 공통 보존적인 OG (Orthologous Group of proteins)를 미토콘드리아 관련 OG와 비관련 OG로 나누어 분석하였다. 62개의 원핵-진핵생물 공통적 COG (Clusters of OG)중 20개가 미토콘드리아 관련 OG였고 이들은 모두 번역관련 OG로 생명현상에서의 단백질의 중요성을 확인할 수 있었다. 세포내 절대기생체인 뇌회백염원충은 비교대상 다른 생물들 모두에 공통적인 미토콘드리아 관련 OG가 전혀 없었다. 뇌회백염원충을 제외한 6개 진핵생물과 원핵생물 63종에 모두 보존적인 미토콘드리아 관련 OG는 17개였다. Phylogenetic tree의 distance 분석을 수행하니 보존적 OG가 원핵생물에서 미토콘드리아 관련 OG와 비관련 OG 등 각각 2개의 그룹으로 나누어 졌고(p<0.001, paired t-test) 진핵생물은 그렇지 않았다(p>0.05, paired t-test). 보존성이 가장 높은 ortholog는 미토콘드리아 관련 OG에서는 COG0048-KOG1750 (ribosomal small subunit S12)이었고, 미토콘드리아 비관련 OG에서는 COG0100-KOG0407 (ribosomal small subunit S11)이었다. 본 연구결과는 진화관계 등의 기초학문적 연구와 치료제 개발 등의 자료가 될 수 있을 것이다.

Phylogenetic Analysis and Rapid Detection of Genus Phellinus using the Nucleotide Sequences of 18S Ribosomal RNA

  • Nam, Byung-Hyouk;Lee, Jae-Yun;Kim, Gi-Young;Jung, Heon-Ho;Park, Hyung-Sik;Kim, Cheng-Yun;Jo, Wol-Soon;Jeong, Soo-Jin;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
    • /
    • 제31권3호
    • /
    • pp.133-138
    • /
    • 2003
  • Analysis of phylogenetic relationship was performed among Phellinus species based on 18S ribosomal subunit sequence data. Twenty-five strains of 19 Phellinus species including P. linteus were examined in this study. Regions of 18S ribosomal subunit were very conserved, but some variable regions between Phellinus species were observed. The species-specific detection primers, modified by 2 or 3 nucleotides in sense primer were designed based on 18S ribosomal DNA(rDNA) sequence data. The 210 by PCR bands were detected with annealing temperature $48^{\circ}C$. The 18S 2F-18S 4R detection primer set distinguished P. linteus from various Phellinus species but some species like P. baumii, P. weirianius, P. rhabarberinus and P. pomaceus also had weak reactivity on this primer set. The 18S 3F-18S 4R primer set distinguished only P. linteus from various Phellinus species, although sensitivity with this primer set was lower than that of 18S 2F-18 4R primer set. These primer sets would be useful for the detection of only P. linteus among unknown Phellinus species rapidly.

Functional Analysis of the Invariant Residue G791 of Escherichia coli 16S rRNA

  • Song, Woo-Seok;Kim, Hong-Man;Kim, Jae-Hong;Sim, Se-Hoon;Ryou, Sang-Mi;Kim, Sang-Goo;Cha, Chang-Jun;Cunningham, Philip R.;Bae, Jee-Hyeon;Lee, Kang-Seok
    • Journal of Microbiology
    • /
    • 제45권5호
    • /
    • pp.418-421
    • /
    • 2007
  • The nucleotide at position 791(G791) of E. coli 16S rRNA was previously identified as an invariant residue for ribosomal function. In order to characterize the functional role of G791, base substitutions were introduced at this position, and mutant ribosomes were analyzed with regard to their protein synthesis ability, via the use of a specialized ribosome system. These ribosomal RNA mutations attenuated the ability of ribosomes to conduct protein synthesis by more than 65%. A transition mutation (G to A) exerted a moderate effect on ribosomal function, whereas a transversion mutation (G to C or U) resulted in a loss of protein synthesis ability of more than 90%. The sucrose gradient profiles of ribosomes and primer extension analysis showed that the loss of protein-synthesis ability of mutant ribosomes harboring a base substitution from G to U at position 791 stems partially from its inability to form 70S ribosomes. These findings show the involvement of the nucleotide at position 791 in the association of ribosomal subunits and protein synthesis steps after 70S formation, as well as the possibility of using 16S rRNA mutated at position 791 for the selection of second-site revertants in order to identify ligands that interact with G791 in protein synthesis.

Mycoplasma pneumoniae의 macrolide 내성과 연관된 유전자 변이의 검출 (Detection of genetic mutations associated with macrolide resistance of Mycoplasma pneumoniae)

  • 오지은;최은화;이환종
    • Clinical and Experimental Pediatrics
    • /
    • 제53권2호
    • /
    • pp.178-183
    • /
    • 2010
  • 목 적 : 최근에 macrolide계 항균제에 내성인 M. pneumoniae 균주가 증가한다는 외국의 보고가 있었으며, 국내에서 수행된 한 연구에서도 M. pneumoniae의 macrolide 내성률을 49% 정도로 보고한 바 있다. 이에, 본 연구는 M. pneumoniae 폐렴으로 진단된 소아의 비인두 흡인물에서 M. pneumoniae의 macrolide 계항균제 내성에 연관된 것으로 알려진 유전자 변이 유무를 확인하고, M. pneumoniae의 macrolide계 항균제에 대한 최소억제농도를 측정하기 위한 기초 연구로 M. pneumoniae 배양법을 구축하고자 시행되었다. 방 법 : 2000년과 2003년 M. pneumoniae 감염의 유행기에 급성 호흡기 증상을 주소로 서울대학교 어린이병원과 분당서울대학교병원에서 치료받은 소아 중 혈청학적 검사와 M. pneumoniae PCR을 통해 M. pneumoniae 폐렴으로 진단받은 환아 62명으로 부터 채취하여 $-80^{\circ}C$에 보관되었던 비인두 흡인물을 대상으로 하였다. M. pneumoniae의 23S rRNA domain V의 peptidyl transferase 부위와 ribosomal protein L4를 M. pneumoniae 특이 PCR로 증폭한 후 염기서열분석을 시행하였다. 염기서열의 분석은 M. pneumoniae 표준 균주와 비교하여, 23S rRNA domain V의 A2063G, A2064G 변이와 ribosomal protein L4의 M144V변이 유무를 확인하였다. 또한, M. pneumoniae 표준 균주와 33개의 비인두흡인물($-80^{\circ}C$에 보관되었던 28검체와 1-2일간 냉장보관되었던 비인두흡인물 5 검체)을 Chanock's glucose 액체배지와 한천배지에 접종하고 $37^{\circ}C$의 5% $CO_2$ 항온기에서 6주간 관찰하여 배양을 확인하였다. 결 과 : 총 62 검체 중 23S rRNA gene에 대한 염기서열분석이 가능했던 61 검체 중 1검체(1.6%)에서 A2064G변이가 관찰되었고, 62 검체의 ribosomal protein L4에 대한 염기서열분석 결과 17검체(27.4%)에서 M144V 아미노산 변이가 확인되었다. M. pneumoniae 배양 결과, 표준 균주는 Chanock's glucose 액체배지와 한천배지 모두에서 배양되었고 2009년에 채취된 5검체 중 2검체에서 배양이 확인되었으나, $-80^{\circ}C$에 보관되었던 28검체는 모두 배양되지 않았다. 결 론 : 본 연구에서 23S rRNA gene의 유전자 변이 빈도는 매우 낮았고, ribosomal protein L4의 M144V 변이는 좀 더 많은 검체에서 확인되었다. Macrolide계 항균제에 내성인 M. pneumoniae의 분포와 M. pneumoniae의 23S rRNA gene과 ribosomal protein L4의 변이에 대한 추가적인 연구들을 통해 M. pneumoniae의 macrolide 항균제에 대한 내성기전을 이해하는데 도움을 줄 수 있을 것으로 생각된다.

Partial Purification of Factors for Differential Transcription of the rrnD Promoters for Ribosomal RNA Synthesis in Streptomyces coelicolor

  • Hahn, Mi-Young;Roe, Jung-Hye
    • Journal of Microbiology
    • /
    • 제45권6호
    • /
    • pp.534-540
    • /
    • 2007
  • The Streptomyces coelicolor A3(2) genome contains six operons (rrnA to F) for ribosomal RNA synthesis. Transcription from rrnD occurs from four promoters (p1 to p4). We found that transcripts from the p1 and p3 promoters were most abundant in vivo in the early exponential phase. However, at later phases of exponential and stationary growth, transcripts from the p1 promoter decreased drastically, with the p3 and p4 transcripts constituting the major forms. Partially purified RNA polymerase supported transcription from the p3 and p4 promoters, whereas pure reconstituted RNA polymerase with core enzyme (E) and the major vegetative sigma factor ${\sigma}^{HrdB}$ ($E{\cdot}{\sigma}^{HrdB}$) did not. In order to assess any potential requirement for additional factor(s) that allow transcription from the p3 and p4 promoters, we fractionated a partially purified RNA polymerase preparation by denaturing gel filtration chromatography. We found that transcription from the p3 and p4 promoters required factor(s) of about 30-35 kDa in addition to RNAP holoenzyme ($E{\cdot}{\sigma}^{HrdB}$). Therefore, transcription from the p3 and p4 promoters, which contain a consensus -10 region but no -35 for ${\sigma}^{HrdB}$ recognition, are likely to be regulated by transcription factor(s) that modulate RNA polymerase holoenzyme activity in S. coelicolor.