Mutation of the HIV-1 5′ss in pNLenv does not relieve the 3′ splice inhibition

<p><b>Copyright information:</b></p><p>Taken from "Splicing of human immunodeficiency virus RNA is position-dependent suggesting sequential removal of introns from the 5′ end"</p><p>Nucleic Acids Research 2005;33(3):825-837.</p><p>Published online 8 Feb 2005</p><p>PMCID:PMC549389.</p><p>© The Author 2005. Published by Oxford University Press. All rights reserved</p> () Schematic depiction of plasmids pNLenvM3 and pNLenvM3BGI. The sequence at the HIV-1 5′ss #4 (wt) and the complementary sequence of the U1 RNA is expanded on top. The three mutations introduced into the 5′ss are depicted below in bold and underlined (M3). () Western blot analysis of HeLa P4 cells transfected with the constructs indicated above each lane. A Tat expression plasmid was cotransfected in all cases. HIV-1 proteins are identified on the right as in . () Northern blot analysis of 10 μg of RNA obtained from the same transfection as in panel B. The upper panel was probed with an LTR-specific probe, the middle panel with a BGI-specific probe, and the bottom panel with a GAPDH-specific probe. The observed RNA species are identified as in ; numbering is according to the drawing in