Sequence-specific activation of the DNA sensor cGAS by Y-form DNA structures as found in primary HIV-1 cDNA.
Cytosolic DNA that emerges during infection with a retrovirus or DNA virus triggers antiviral type I interferon responses. So far, only double-stranded DNA (dsDNA) over 40 base pairs (bp) in length has been considered immunostimulatory. Here we found that unpaired DNA nucleotides flanking short base-paired DNA stretches, as in stem-loop structures of single-stranded DNA (ssDNA) derived from human immunodeficiency virus type 1 (HIV-1), activated the type I interferon-inducing DNA sensor cGAS in a sequence-dependent manner. DNA structures containing unpaired guanosines flanking short (12- to 20-bp) dsDNA (Y-form DNA) were highly stimulatory and specifically enhanced the enzymatic activity of cGAS. Furthermore, we found that primary HIV-1 reverse transcripts represented the predominant viral cytosolic DNA species during early infection of macrophages and that these ssDNAs were highly immunostimulatory. Collectively, our study identifies unpaired guanosines in Y-form DNA as a highly active, minimal cGAS recognition motif that enables detection of HIV-1 ssDNA.
Institute of Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, Bonn, Germany.
Anna-Maria
AM
Herzner
Institute of Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, Bonn, Germany.
Cristina Amparo
CA
Hagmann
Institute of Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, Bonn, Germany.
Marion
M
Goldeck
Institute of Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, Bonn, Germany.
Steven
S
Wolter
Department of Obstetrics and Gynecology, Center for Integrated Oncology, University of Bonn, Bonn, Germany.
Kirsten
K
Kübler
Institute of Clinical and Molecular Virology, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany.
Sabine
S
Wittmann
Institute of Clinical and Molecular Virology, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany.
Thomas
T
Gramberg
Department Biochemistry, Gene Center, Ludwig-Maximilians University, Munich, Germany.
Liudmila
L
Andreeva
Department Biochemistry, Gene Center, Ludwig-Maximilians University, Munich, Germany.
Karl-Peter
KP
Hopfner
Institute of Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, Bonn, Germany.
Christina
C
Mertens
Institute of Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, Bonn, Germany.|German Center of Infectious Disease, Cologne-Bonn, Germany.
Thomas
T
Zillinger
School of Life Sciences, University of Science and Technology of China, Hefei, China.
Tengchuan
T
Jin
Department of Pathology, Case Western Reserve University, Cleveland, Ohio, USA.
Tsan Sam
http://orcid.org/0000-0001-9688-475X
TS
Xiao
Institute of Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, Bonn, Germany.
Eva
E
Bartok
Institute of Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, Bonn, Germany.
Christoph
C
Coch
LIMES Institute, Chemical Biology, University of Bonn, Bonn, Germany.
Damian
D
Ackermann
Institute of Molecular Medicine, University Hospital, University of Bonn, Bonn, Germany.
Veit
V
Hornung
Institute of Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, Bonn, Germany.
Janos
J
Ludwig
Institute of Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, Bonn, Germany.|German Center of Infectious Disease, Cologne-Bonn, Germany.
Winfried
W
Barchet
Institute of Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, Bonn, Germany.
Gunther
G
Hartmann
Institute of Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, Bonn, Germany.
Martin
http://orcid.org/0000-0003-3671-7639
M
Schlee
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D004279:DNA, Viral / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D057809:HEK293 Cells; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D007114:Immunization; D016898:Interferon-alpha / Q000276:immunology*; D051379:Mice; D009713:Nucleotidyltransferases / Q000235:genetics*; D000099235:Cyclic Guanosine Monophosphate-Adenosine Monophosphate Synthase
D018076:DNA, Complementary; D004279:DNA, Viral; D016898:Interferon-alpha; D009713:Nucleotidyltransferases; C578391:cGAS protein, mouse; D000099235:Cyclic Guanosine Monophosphate-Adenosine Monophosphate Synthase