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Journal of Virology, November 2001, p. 10670-10682, Vol. 75, No. 22
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.22.10670-10682.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

A New RNA Element Located in the Coding Region of a Murine Endogenous Retrovirus Can Functionally Replace the Rev/Rev-Responsive Element System in Human Immunodeficiency Virus Type 1 Gag Expression

Harald Wodrich,^1, Jens Bohne,^1,2 Ellen Gumz,¹ Reinhold Welker,^1, and Hans-Georg Kräusslich^1,2,*

Heinrich-Pette-Institut, D-20251 Hamburg,¹ and Abteilung Virologie, Universität Heidelberg, D-69120 Heidelberg,² Germany

Received 8 March 2001/Accepted 7 August 2001

Nuclear export of incompletely spliced RNAs is a prerequisite for retroviral replication. Complex retroviruses like human immunodeficiency virus (HIV) encode a viral transport factor (Rev), which binds to its target sequence on the RNA genome and directs it into the Crm-1-mediated export pathway. Other retroviruses, like Mason-Pfizer monkey virus, contain cis-acting constitutive RNA transport elements (CTE) which achieve nuclear export of intron-containing RNA via cellular transport factors. Here, we describe the identification and characterization of a novel cis-acting orientation-dependent RNA expression element in the coding region of the murine intracisternal A-type particle (IAP) MIA14. This IAP expression element (IAPE) can functionally replace the Rev system in the expression of HIV-1 Gag proteins but functions independently of Crm-1. The presence of this element is needed for the expression of the IAP Gag proteins, indicating its biological significance. The IAPE can be functionally replaced by placing a CTE on the MIA14 RNA, further supporting its role in mRNA export. Northern blot analysis revealed that total RNA, as well as cytoplasmic RNA, was increased when the element was present. The element was mapped to a predicted stem-loop structure in the 3' part of the pol open reading frame. There was no overall homology between the IAPE and the CTE, but there was complete sequence identity between short putative single-stranded loops. Deletion of these loops from the IAPE severely reduced Rev-independent Gag expression.

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^* Corresponding author. Mailing address: Abteilung Virologie, Universität Heidelberg, Im Neuenheimer Feld 324, D-69120 Heidelberg, Germany. Phone: 49 6221 56-5001. Fax: 49 6221 56-5003. E-mail: Hans-Georg_Kraeusslich{at}med.uni-heidelberg.de.

Present address: The Scripps Research Institute, Department of Cell Biology, La Jolla, CA 92037.

Present address: Bayer AG, Virologie, D-42069 Wuppertal, Germany.

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Journal of Virology, November 2001, p. 10670-10682, Vol. 75, No. 22
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.22.10670-10682.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

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