| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Animal: RNA Viruses |
Department of Microbiology and Immunology1 and Department of Medicine2, The Milton S. Hershey Medical Center, Pennsylvania State University College of Medicine, PO Box 850, Hershey, PA 17033-0850, USA
Author for correspondence: Michael Katzman (at the Department of Medicine). Fax +1 717 531 4633. e-mail mkatzman{at}psu.edu
Many reports describe the characteristics of susceptible viral DNA substrates to various retroviral integrases during in vitro reactions in which manganese serves as the divalent cation cofactor for site-specific nicking. However, manganese is known to alter the specificity of some endonucleases and magnesium may be the divalent cation used during retroviral integration in vivo. To address these concerns, we identified conditions under which the integrases of human immunodeficiency virus type 1 and visna virus were optimally active with magnesium (the first time such activity was shown for visna virus integrase) and used these conditions to test the susceptibility of a series of oligodeoxynucleotide substrates. The data show that two base pairs immediately internal to the conserved CA dinucleotide near the termini of retroviral DNA are selectively recognized by the two integrases and that the final six base pairs of viral DNA contain sufficient sequence information for specific recognition and cleavage by each enzyme. The results validate the importance of the subterminal viral DNA positions even in the presence of magnesium and identify viral DNA positions that functionally interact with integrase. The data obtained under magnesium-dependent conditions, which were obtained with substrates containing single and multiple base-pair substitutions and two different retroviral integrases, are consistent with those previously obtained with manganese. Thus, the large body of manganese-dependent data identifying terminal viral DNA positions that are important in substrate recognition by various integrases likely reflects interactions that are biologically relevant.
This article has been cited by other articles:
|
|
 |
|
  P. Cherepanov LEDGF/p75 interacts with divergent lentiviral integrases and modulates their enzymatic activity in vitro Nucleic Acids Res., January 12, 2007; 35(1): 113 - 124. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. M. Konsavage Jr., S. Burkholder, M. Sudol, A. L. Harper, and M. Katzman A Substitution in Rous Sarcoma Virus Integrase That Separates Its Two Biologically Relevant Enzymatic Activities J. Virol., April 15, 2005; 79(8): 4691 - 4699. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. L. Harper, M. Sudol, and M. Katzman An Amino Acid in the Central Catalytic Domain of Three Retroviral Integrases That Affects Target Site Selection in Nonviral DNA J. Virol., March 15, 2003; 77(6): 3838 - 3845. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Brin and J. Leis HIV-1 Integrase Interaction with U3 and U5 Terminal Sequences in Vitro Defined Using Substrates with Random Sequences J. Biol. Chem., May 17, 2002; 277(21): 18357 - 18364. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. L. Harper, L. M. Skinner, M. Sudol, and M. Katzman Use of Patient-Derived Human Immunodeficiency Virus Type 1 Integrases To Identify a Protein Residue That Affects Target Site Selection J. Virol., August 15, 2001; 75(16): 7756 - 7762. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. M. Skinner, M. Sudol, A. L. Harper, and M. Katzman Nucleophile Selection for the Endonuclease Activities of Human, Ovine, and Avian Retroviral Integrases J. Biol. Chem., January 5, 2001; 276(1): 114 - 124. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| INT J SYST EVOL MICROBIOL | MICROBIOLOGY | J GEN VIROL |
| J MED MICROBIOL | ALL SGM JOURNALS | |
