BACTERIOPHAGE-T4 DDA HELICASE TRANSLOCATES IN A UNIDIRECTIONAL FASHION ON SINGLE-STRANDED-DNA

被引:55
作者
RANEY, KD [1 ]
BENKOVIC, SJ [1 ]
机构
[1] PENN STATE UNIV,DEPT CHEM,DAVEY LAB 152,UNIVERSITY PK,PA 16802
关键词
D O I
10.1074/jbc.270.38.22236
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The T4 bacteriophage Dda helicase is believed to be involved in early events in T4 DNA replication and has been shown to stimulate genetic recombination processes in vitro. Dda unwinds double-stranded DNA with 5' to 3' polarity but its ability to translocate on DNA has not been established. The DNA stimulated ATPase activity of Dda helicase has been used to probe translocation on single-strand DNA (ssDNA). Dda exhibits higher ATPase activity in the presence of poly(dT) than oligo(dT)(16), indicating that Dda translocates on ssDNA. Oligonucleotides containing biotin/streptavidin blocks on the 5' or 3' end were used to probe directionality of translocation. The K-act (K-m for DNA) for Dda ATPase activity was reduced in the presence of a streptavidin block. on the 3' end, whereas a streptavidin block on the 5' end had only a small effect on the steady-state ATPase parameters. These results suggest that Dda translocates unidirectionally in a 5' to 3' manner and upon encountering the block remains bound 60 the oligonucleotide rather than sliding off the 3' end. The direction of translocation on ssDNA is consistent with the direction in which Dda unwinds duplex DNA and is not dependent on duplex structure.
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页码:22236 / 22242
页数:7
相关论文
共 32 条
[1]   ESCHERICHIA-COLI REP HELICASE UNWINDS DNA BY AN ACTIVE MECHANISM [J].
AMARATUNGA, M ;
LOHMAN, TM .
BIOCHEMISTRY, 1993, 32 (27) :6815-6820
[2]  
BARRY J, 1994, J BIOL CHEM, V269, P33063
[3]   ESCHERICHIA-COLI REPLICATION TERMINATOR PROTEIN IMPEDES SIMIAN VIRUS-40 (SV40) DNA-REPLICATION FORK MOVEMENT AND SV40 LARGE TUMOR-ANTIGEN HELICASE ACTIVITY INVITRO AT A PROKARYOTIC TERMINUS SEQUENCE [J].
BEDROSIAN, CL ;
BASTIA, D .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (07) :2618-2622
[4]  
CHAMBERLIN MJ, 1965, FED PROC, V24, P1446
[5]  
CHAO KL, 1990, J BIOL CHEM, V265, P1067
[6]   AFFINITY PURIFICATION OF BACTERIOPHAGE-T4 PROTEINS ESSENTIAL FOR DNA-REPLICATION AND GENETIC-RECOMBINATION [J].
FORMOSA, T ;
BURKE, RL ;
ALBERTS, BM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1983, 80 (09) :2442-2446
[7]   DNA HELICASE REQUIREMENTS FOR DNA-REPLICATION DURING BACTERIOPHAGE-T4 INFECTION [J].
GAUSS, P ;
PARK, K ;
SPENCER, TE ;
HACKER, KJ .
JOURNAL OF BACTERIOLOGY, 1994, 176 (06) :1667-1672
[8]   A PHYSICAL MODEL FOR THE TRANSLOCATION AND HELICASE ACTIVITIES OF ESCHERICHIA-COLI TRANSCRIPTION TERMINATION PROTEIN-RHO [J].
GEISELMANN, J ;
WANG, Y ;
SEIFRIED, SE ;
VONHIPPEL, PH .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (16) :7754-7758
[9]  
HACKER KJ, 1992, J BIOL CHEM, V267, P20674
[10]  
JONGENEEL CV, 1984, J BIOL CHEM, V259, P2925