Nullbasic

To investigate the molecular effects of transdominant Tat mutants, a novel mutant termed Nullbasic was created.

PLoS ONE Alerts: New Articles Luke W. Meredith et al. 2009

Sequestration of cofactors by transdominant Tat mutants may be mediated by an intact activation domain, but due to the disrupted basic domain, Tat mutants may be incapable of recruiting the cofactors (e.g., cyclin T1) to the site of transactivation Here we describe for the first time the transdominant effects of a two-exon, 101 amino-acid Tat mutant called Nullbasic, which has the entire basic domain (amino acids 49-57) replaced with glycine/alanine residues.

PLoS ONE Alerts: New Articles Luke W. Meredith et al. 2009

Here we describe for the first time a full-length, transdominant two-exon Tat mutant, termed Nullbasic

PLoS ONE Alerts: New Articles Luke W. Meredith et al. 2009

Queensland, Australia Herein we describe a mutant of the two-exon HIV-1 Tat protein, termed Nullbasic, that potently inhibits multiple steps of the HIV-1 replication cycle.

PLoS ONE Alerts: New Articles Luke W. Meredith et al. 2009

To further examine the production of HIV-1 viral proteins, western blotting was performed on β-galactosidase-equalized lysates of cells co-expressing pGCH provirus and either Nullbasic or Tat-FLAG.

PLoS ONE Alerts: New Articles Luke W. Meredith et al. 2009

Virions were generated in HEK293T cells co-transfected with 1: 4 molar ratios of pGCH provirus to wild type Tat-FLAG, Nullbasic or empty vector ( "No Tat") plasmids.

PLoS ONE Alerts: New Articles Luke W. Meredith et al. 2009

(A) The infectivity of HIV-1 produced by HEK293T cells expressing pGCH provirus co-transfected with empty vector (pcDNA3. 1+) or co-expressing increasing amounts of Nullbasic or Tat-FLAG (at 1: 2 and 1: 4 molar ratios) were determined by the MAGI assay.

PLoS ONE Alerts: New Articles Luke W. Meredith et al. 2009

HIV-1 mRNAs isolated from HEK293T cells co-expressing pGCH provirus and either Tat-FLAG or Nullbasic were detected with a single probe capable of binding to the unspliced, singly-spliced and multiply-spliced classes of HIV-1 transcripts.

PLoS ONE Alerts: New Articles Luke W. Meredith et al. 2009

The concentration of RT was similarly decreased, indicating that Nullbasic expression suppressed overall virus production.

PLoS ONE Alerts: New Articles Luke W. Meredith et al. 2009

Env and Nullbasic were detected by immunoblotting with anti-gp120 and anti-FLAG antibodies, respectively.

PLoS ONE Alerts: New Articles Luke W. Meredith et al. 2009