INVITRO ASSEMBLY OF RELAXOSOMES AT THE TRANSFER ORIGIN OF PLASMID-RP4

被引：125

作者：

PANSEGRAU, W ^{[1
]}

BALZER, D ^{[1
]}

KRUFT, V ^{[1
]}

LURZ, R ^{[1
]}

LANKA, E ^{[1
]}

机构：

[1] MAX PLANCK INST MOLEC GENET,SCHUSTER ABT,IHNESTR 73,W-1000 BERLIN 33,GERMANY

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 1990年 / 87卷 / 17期

关键词：

bacterial conjugation; electron microscopy; initiation of transfer DNA replication; protein-DNA interaction;

D O I：

10.1073/pnas.87.17.6555

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

During initiation of conjugative transfer of DNA containing the transfer origin (oriT) of the promiscuous plasmid RP4, the proteins TraI, TraJ, and TraH interact and assemble a specialized nucleoprotein complex (the relaxosome) at oriT. The structure can be visualized on electron micrographs. Site- and strand-specific nicking at the transfer origin in vitro is dependent on the proteins TraI and TraJ and on Mg2+ ions. Substrate specificity is directed exclusively towards the cognate transfer origin: the RP4-specified TraJ protein cannot recognize the closely related oriT of plasmid R751. After nicking, TraI protein remains attached to the 5'-terminal 2'-deoxycytidyl residue at the nic site [Pansegrau, W., Ziegelin, G. and Lanka, E. (1990) J. Biol. Chem. 265, 10637-10644]. Nicking and relaxosome formation require supercoiled DNA. Thus, a complicated structure involving multiple plasmid-specified proteins and a defined region of DNA must be formed at the transfer origin to prepare the plasmid for generating the single strand to be transferred.

引用

页码：6555 / 6559

页数：5

共 15 条

[1] MOLECULAR-CLONING OF THE PLASMID RP4 PRIMASE REGION IN A MULTI-HOST-RANGE TACP EXPRESSION VECTOR [J].

FURSTE, JP ;

PANSEGRAU, W ;

FRANK, R ;

BLOCKER, H ;

SCHOLZ, P ;

BAGDASARIAN, M ;

LANKA, E .

GENE, 1986, 48 (01) :119-131

[2] CONJUGATIVE TRANSFER OF PROMISCUOUS INCP PLASMIDS - INTERACTION OF PLASMID-ENCODED PRODUCTS WITH THE TRANSFER ORIGIN [J].

FURSTE, JP ;

PANSEGRAU, W ;

ZIEGELIN, G ;

KROGER, M ;

LANKA, E .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (06) :1771-1775

[3] DNA-PROTEIN RELAXATION COMPLEX OF THE PLASMID RK2 - LOCATION OF THE SITE-SPECIFIC NICK IN THE REGION OF THE PROPOSED ORIGIN OF TRANSFER [J].

GUINEY, DG ;

HELINSKI, DR .

MOLECULAR & GENERAL GENETICS, 1979, 176 (02) :183-189

[4]

GUINEY DG, 1989, PROMISCUOUS PLASMIDS, P27

[5] STUDIES ON TRANSFORMATION OF ESCHERICHIA-COLI WITH PLASMIDS [J].

HANAHAN, D .

JOURNAL OF MOLECULAR BIOLOGY, 1983, 166 (04) :557-580

[6] BACTERIAL CONJUGATIVE PLASMIDS MOBILIZE DNA TRANSFER BETWEEN BACTERIA AND YEAST [J].

HEINEMANN, JA ;

SPRAGUE, GF .

NATURE, 1989, 340 (6230) :205-209

[7]

HEWICK RM, 1981, J BIOL CHEM, V256, P7990

[8]

Maniatis T., 1982, MOL CLONING

[9]

PANSEGRAU W, 1990, J BIOL CHEM, V265, P10637

[10] THE ORIGIN OF CONJUGATIVE INCP PLASMID TRANSFER - INTERACTION WITH PLASMID-ENCODED PRODUCTS AND THE NUCLEOTIDE-SEQUENCE AT THE RELAXATION SITE [J].

PANSEGRAU, W ;

ZIEGELIN, G ;

LANKA, E .

BIOCHIMICA ET BIOPHYSICA ACTA, 1988, 951 (2-3) :365-374

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