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Akusjärvi, Göran
Alternative names
Publications (10 of 93) Show all publications
Kamel, W. & Akusjärvi, G. (2017). An Ago2-associated capped transcriptional start site small RNA suppresses adenovirus DNA replication. RNA: A publication of the RNA Society, 23(11), 1700-1711
Open this publication in new window or tab >>An Ago2-associated capped transcriptional start site small RNA suppresses adenovirus DNA replication
2017 (English)In: RNA: A publication of the RNA Society, ISSN 1355-8382, E-ISSN 1469-9001, Vol. 23, no 11, p. 1700-1711Article in journal (Refereed) Published
Abstract [en]

Here we show that the adenovirus major late promoter produces a 31-nucleotide transcriptional start site small RNA (MLP-TSS-sRNA) that retains the 7-methylguanosine (m7G)-cap and is incorporated onto Ago2-containing RNA-induced silencing complexes (RISC) in human adenovirus-37 infected cells. RNA polymerase II CLIP (UV-cross linking immunoprecipitation) experiments suggest that the MLP-TSS-sRNA is produced by promoter proximal stalling/termination of RNA polymerase II transcription at the site of the small RNA 3' end. The MLP-TSS-sRNA is highly stable in cells and functionally active, down-regulating complementary targets in a sequence and dose-dependent manner. The MLP-TSS-sRNA is transcribed from the opposite strand to the adenoviral DNA polymerase and preterminal protein mRNAs, two essential viral replication proteins. We show that the MLP-TSS-sRNA act in trans to reduce DNA polymerase and preterminal protein mRNA expression. As a consequence of this, the MLP-TSS-sRNA has an inhibitory effect on the efficiency of viral DNA replication. Collectively, our results suggest that this novel sRNA may serve a regulatory function controlling viral genome replication during a lytic and/or persistent adenovirus infection in its natural host.

Keyword
adenovirus, Ago2, m7G-cap, small RNA
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
urn:nbn:se:uu:diva-339747 (URN)10.1261/rna.061291.117 (DOI)000412996100009 ()28839112 (PubMedID)
Funder
Swedish Cancer Society, 130469Swedish Research Council, 2006-5038-36531-16
Available from: 2018-01-25 Created: 2018-01-25 Last updated: 2018-01-25Bibliographically approved
Assadian, F., Kamel, W., Laurell, G., Svensson, C., Punga, T. & Akusjärvi, G. (2017). Expression profile of Epstein-Barr virus and human adenovirus small RNAs in tonsillar B and T lymphocytes. PLoS ONE, 12(5), Article ID e0177275.
Open this publication in new window or tab >>Expression profile of Epstein-Barr virus and human adenovirus small RNAs in tonsillar B and T lymphocytes
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2017 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 12, no 5, article id e0177275Article in journal (Refereed) Published
Abstract [en]

We have used high-throughput small RNA sequencing to characterize viral small RNA expression in purified tonsillar B and T lymphocytes isolated from patients tested positive for Epstein-Barr virus (EBV) or human adenovirus (HAdV) infections, respectively. In the small set of patients analyzed, the expression profile of EBV and HAdV miRNAs could not distinguish between patients diagnosed with tonsillar hypertrophy or chronic/recurrent tonsillitis. The EBV miR-BART expression profile among the patients diagnosed with tonsillar diseases resembles most closely the pattern seen in EBV+ tumors (Latency II/I). The miRBARTs that appear to be absent in normal EBV infected cells are essentially all detectable in the diseased tonsillar B lymphocytes. In the EBV+ B cells we detected 44 EBV miRBARTs derived from the proposed BART precursor hairpins whereof five are not annotated in miRBase v21. One previously undetected miRNA, BART16b-5p, originates from the miR-BART16 precursor hairpin as an alternative 5 A miR-BART16 located precisely upstream of the annotated miR-BART16-5p. Further, our analysis revealed an extensive sequence variation among the EBV miRNAs with isomiRs having a constant 5 A end but alternative 3 A ends. A range of small RNAs was also detected from the terminal stem of the EBER RNAs and the 3 A part of v-snoRNA1. During a lytic HAdV infection in established cell lines the terminal stem of the viral non-coding VA RNAs are processed to highly abundant viral miRNAs (mivaRNAs). In contrast, mivaRNA expression in HAdV positive tonsillar T lymphocytes was very low. The small RNA profile further showed that the 5 A mivaRNA from VA RNAI and the 3 A mivaRNA from VA RNAII were as predicted, whereas the 3 A mivaRNA from VA RNAI showed an aberrant processing upstream of the expected Dicer cleavage site.

Place, publisher, year, edition, pages
PUBLIC LIBRARY SCIENCE, 2017
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-326234 (URN)10.1371/journal.pone.0177275 (DOI)000402062800013 ()28542273 (PubMedID)
Funder
Swedish Cancer Society, 120678, 130469
Available from: 2017-07-06 Created: 2017-07-06 Last updated: 2017-11-29Bibliographically approved
Lan, S., Kamel, W., Punga, T. & Akusjärvi, G. (2017). The adenovirus L4-22K protein regulates transcription and RNA splicing via a sequence-specific single-stranded RNA binding. Nucleic Acids Research, 45(4), 1731-1742
Open this publication in new window or tab >>The adenovirus L4-22K protein regulates transcription and RNA splicing via a sequence-specific single-stranded RNA binding
2017 (English)In: Nucleic Acids Research, ISSN 0305-1048, E-ISSN 1362-4962, Vol. 45, no 4, p. 1731-1742Article in journal (Refereed) Published
Abstract [en]

The adenovirus L4-22K protein both activates and suppresses transcription from the adenovirus major late promoter (MLP) by binding to DNA elements located downstream of the MLP transcriptional start site: the so-called DE element ( positive) and the R1 region ( negative). Here we show that L4-22K preferentially binds to the RNA form of the R1 region, both to the double-stranded RNA and the single-stranded RNA of the same polarity as the nascent MLP transcript. Further, L4-22K binds to a 5'-CAAA-3' motif in the single-stranded RNA, which is identical to the sequence motif characterized for L4-22K DNA binding. L4-22K binding to single-stranded RNA results in an enhancement of U1 snRNA recruitment to the major late first leader 5' splice site. This increase in U1 snRNA binding results in a suppression of MLP transcription and a concurrent stimulation of major late first intron splicing.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-318946 (URN)10.1093/nar/gkw1145 (DOI)000396055400023 ()27899607 (PubMedID)
Funder
Swedish Cancer Society, 120678, 130410Swedish Research Council, 2006-5038-36531-16
Available from: 2017-04-03 Created: 2017-04-03 Last updated: 2017-11-29Bibliographically approved
Assadian, F., Sandström, K., Bondeson, K., Laurell, G., Lidian, A., Svensson, C., . . . Punga, T. (2016). Distribution and Molecular Characterization of Human Adenovirus and Epstein-Barr Virus Infections in Tonsillar Lymphocytes Isolated from Patients Diagnosed with Tonsillar Diseases. PLoS ONE, 11(5), Article ID e0154814.
Open this publication in new window or tab >>Distribution and Molecular Characterization of Human Adenovirus and Epstein-Barr Virus Infections in Tonsillar Lymphocytes Isolated from Patients Diagnosed with Tonsillar Diseases
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2016 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 11, no 5, article id e0154814Article in journal (Refereed) Published
Abstract [en]

Surgically removed palatine tonsils provide a conveniently accessible source of T and B lymphocytes to study the interplay between foreign pathogens and the host immune system. In this study we have characterised the distribution of human adenovirus (HAdV), Epstein-Barr virus (EBV) and human cytomegalovirus (HCMV) in purified tonsillar T and B cell-enriched fractions isolated from three patient age groups diagnosed with tonsillar hypertrophy and chronic/recurrent tonsillitis. HAdV DNA was detected in 93 out of 111 patients (84%), while EBV DNA was detected in 58 patients (52%). The most abundant adenovirus type was HAdV-5 (68%). None of the patients were positive for HCMV. Furthermore, 43 patients (39%) showed a co-infection of HAdV and EBV. The majority of young patients diagnosed with tonsillar hypertrophy were positive for HAdV, whereas all adult patients diagnosed with chronic/recurrent tonsillitis were positive for either HAdV or EBV. Most of the tonsils from patients diagnosed with either tonsillar hypertrophy or chronic/recurrent tonsillitis showed a higher HAdV DNA copy number in T compared to B cell-enriched fraction. Interestingly, in the majority of the tonsils from patients with chronic/recurrent tonsillitis HAdV DNA was detected in T cells only, whereas hypertrophic tonsils demonstrated HAdV DNA in both T and B cell-enriched fractions. In contrast, the majority of EBV positive tonsils revealed a preference for EBV DNA accumulation in the B cell-enriched fraction compared to T cell fraction irrespective of the patients' age.

National Category
Surgery Otorhinolaryngology Cancer and Oncology
Identifiers
urn:nbn:se:uu:diva-297795 (URN)10.1371/journal.pone.0154814 (DOI)000375674800050 ()27136093 (PubMedID)
Funder
Swedish Cancer Society, 12 0504Swedish Cancer Society, 13 0469Swedish Research Council, K2012-99X-21959-01-3Swedish Research Council, 2006-5038-36531-16Åke Wiberg Foundation
Note

De 2 sista författarna delar sistaförfattarskapet.

Available from: 2016-06-28 Created: 2016-06-28 Last updated: 2017-11-28Bibliographically approved
Wu, C., Cao, X., Yu, D., Huijbers, E. J. M., Essand, M., Akusjärvi, G., . . . Svensson, C. (2016). HAdV-2-suppressed growth of SV40 T antigen-transformed mouse mammary epithelial cell-induced tumours in SCID mice. Virology, 489, 44-50
Open this publication in new window or tab >>HAdV-2-suppressed growth of SV40 T antigen-transformed mouse mammary epithelial cell-induced tumours in SCID mice
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2016 (English)In: Virology, ISSN 0042-6822, E-ISSN 1096-0341, Vol. 489, p. 44-50Article in journal (Refereed) Published
Abstract [en]

Human adenovirus (HAdV) vectors are promising tools for cancer therapy, but the shortage of efficient animal models for productive HAdV infections has restricted the evaluation of systemic effects to mainly immunodeficient mice. Previously, we reported a highly efficient replication of HAdV-2 in a non-tumorigenic mouse mammary epithelial cell line, NMuMG. Here we show that HAdV-2 gene expression and progeny formation in NMuMG cells transformed with the SV40 T antigen (NMuMG-T cells) were as efficient as in the parental NMuMG cells. Injection of HAdV-2 into tumours established by NMuMG-T in SCID mice caused reduced tumour growth and signs of intratumoural lesions. HAdV-2 replicated within the NMuMG-T-established tumours, but not in interspersed host-derived tissues within the tumours. The specific infection of NMuMG-T-derived tumours was verified by the lack of viral DNA in kidney, lung or spleen although low levels of viral DNA was occasionally found in liver.

Keyword
Oncolytic adenovirus, Murine cells, Anti-tumour activity, Permissive infection, Viral replication
National Category
Infectious Medicine
Identifiers
urn:nbn:se:uu:diva-282389 (URN)10.1016/j.virol.2015.11.031 (DOI)000370892100005 ()26707269 (PubMedID)
Funder
Swedish Cancer Society
Available from: 2016-04-05 Created: 2016-04-05 Last updated: 2017-11-30Bibliographically approved
Lan, S., Östberg, S., Punga, T. & Akusjärvi, G. (2015). A suppressive effect of Sp1 recruitment to the first leader 5' splice site region on L4-22K-mediated activation of the adenovirus major late promoter. Virus Research, 210, 133-140
Open this publication in new window or tab >>A suppressive effect of Sp1 recruitment to the first leader 5' splice site region on L4-22K-mediated activation of the adenovirus major late promoter
2015 (English)In: Virus Research, ISSN 0168-1702, E-ISSN 1872-7492, Vol. 210, p. 133-140Article in journal (Refereed) Published
Abstract [en]

Transcription from the adenovirus major late promoter (MLP) requires binding of late phase-specific factors to the so-called DE element located approximately 100 base pairs downstream of the MLP transcriptional start site. The adenovirus L4-22K protein binds to the DE element and stimulates transcription from the MLP via a DE sequence-dependent mechanism. Here we use a transient expression approach to show that L4-22K binds to an additional site downstream of the MLP start site, the so-called R1 region, which includes the major late first leader 5' splice site. Binding of L4-22K to R1 has a suppressive effect on MLP transcription. L4-22K binds to the distal part of R1 and stimulates the recruitment of Sp1 and other cellular factors to a site overlapping the first leader 5' splice site. Binding of Sp1 to the 5' splice site region had an inhibitory effect on L4-22K-activated MLP transcription.

National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
urn:nbn:se:uu:diva-275814 (URN)10.1016/j.virusres.2015.07.026 (DOI)000365455500020 ()26247419 (PubMedID)
Funder
Swedish Cancer Society, 12 0504Swedish Research Council, K2012-99X-21959-01-3 2006-5038-36531-16
Available from: 2016-02-07 Created: 2016-02-07 Last updated: 2017-11-30Bibliographically approved
Inturi, R., Kamel, W., Akusjärvi, G. & Punga, T. (2015). Complementation of the human adenovirus type 5 VA RNAI defect by the Vaccinia virus E3L protein and serotype-specific VA RNAIs. Virology, 485, 25-35
Open this publication in new window or tab >>Complementation of the human adenovirus type 5 VA RNAI defect by the Vaccinia virus E3L protein and serotype-specific VA RNAIs
2015 (English)In: Virology, ISSN 0042-6822, E-ISSN 1096-0341, Vol. 485, p. 25-35Article in journal (Refereed) Published
Abstract [en]

Human adenoviruses (HAdVs) encode for multifunctional non-coding virus-associated (VA) RNAs, which function as powerful suppressors of the cellular interferon (IFN) and RNA interference (RNAi) systems. In this study we tested the ability of various plant and animal virus encoded RNAi and IFN suppressor proteins to functionally substitute for the HAdV-5 VA RNAI. Our results revealed that only the Vaccinia virus (VACV) E3L protein was able to substitute for the HAdV-5 VA RNAI functions in virus-infected cells. Interestingly, the E3L protein rescues the translational defect but does not stimulate viral capsid mRNA accumulation observed with VA RNA. We further show that the E3L C-terminal region containing the dsRNA-binding domain is needed to enhance VA RNAI mutant virus replication. Additionally, we show that the HAdV-4 and HAdV-37 VA RNAI are more effective than the HAdV-5 VA RNAI in rescuing virus replication.

Keyword
Adenovirus, Vaccinia virus, interferon, PKR, RNAi
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Research subject
Medical Virology
Identifiers
urn:nbn:se:uu:diva-258879 (URN)10.1016/j.virol.2015.07.002 (DOI)000363993100003 ()
Funder
Swedish Cancer Society, 12 0504, 13 0469Swedish Research Council, K2012–999X-21959-01-301–3Åke Wiberg Foundation, M14-01555
Available from: 2015-07-21 Created: 2015-07-21 Last updated: 2017-12-04Bibliographically approved
Assadian, F., Sandström, K., Laurell, G., Svensson, C., Akusjärvi, G. & Punga, T. (2015). Efficient Isolation Protocol for B and T Lymphocytes from Human Palatine Tonsils. Journal of Visualized Experiments, 105, Article ID e53374.
Open this publication in new window or tab >>Efficient Isolation Protocol for B and T Lymphocytes from Human Palatine Tonsils
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2015 (English)In: Journal of Visualized Experiments, ISSN 1940-087X, E-ISSN 1940-087X, Vol. 105, article id e53374Article in journal (Refereed) Published
Abstract [en]

Palatine tonsils are a rich source of B and T lymphocytes. Here we provide an easy, efficient and rapid protocol to isolate B and T lymphocytes from human palatine tonsils. The method described has been specifically adapted for studies of the viral etiology of tonsil inflammation known as tonsillitis.

National Category
Microbiology in the medical area
Identifiers
urn:nbn:se:uu:diva-274679 (URN)10.3791/53374 (DOI)000368573900061 ()
Funder
Swedish Cancer Society, 11 0253, 13 0469Swedish Research Council, K2012-99X-21959-01-3 2006-5038-36531-16
Available from: 2016-01-25 Created: 2016-01-25 Last updated: 2018-01-10Bibliographically approved
Pickl, J. M., Kamel, W., Ciftci, S., Punga, T. & Akusjärvi, G. (2015). Opposite expression of CYP51A1 and its natural antisense transcript AluCYP51A1 in adenovirus type 37 infected retinal pigmented epithelial cells. FEBS Letters, 589(12), 1383-1388
Open this publication in new window or tab >>Opposite expression of CYP51A1 and its natural antisense transcript AluCYP51A1 in adenovirus type 37 infected retinal pigmented epithelial cells
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2015 (English)In: FEBS Letters, ISSN 0014-5793, E-ISSN 1873-3468, Vol. 589, no 12, p. 1383-1388Article in journal (Refereed) Published
Abstract [en]

Cytochrome P450 family member CYP51A1 is a key enzyme in cholesterol biosynthesis whose deregulation is implicated in numerous diseases, including retinal degeneration. Here we describe that HAdV-37 infection leads to downregulation of CYP51A1 expression and overexpression of its antisense non-coding Alu element (AluCYP51A1) in retinal pigment epithelium (RPE) cells. This change in gene expression is associated with a reversed accumulation of a positive histone mark at the CYP51A1 and AluCYP51A1 promoters. Further, transient AluCYP51A1 RNA overexpression correlates with reduced CYP51A1 mRNA accumulation. Collectively, our data suggest that AluCYP51A1 might control CYP51A1 gene expression in HAdV-37-infected RPE cells. (C) 2015 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Keyword
Adenovirus, Human adenovirus type 37, Retinal pigment epithelium, Alu element, CYP51A1
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
urn:nbn:se:uu:diva-255066 (URN)10.1016/j.febslet.2015.04.018 (DOI)000354290500014 ()25907535 (PubMedID)
Funder
Swedish Research Council, K2012-99X-21959-01-3Swedish Research Council, 2006-5038-36531-16
Available from: 2015-06-22 Created: 2015-06-12 Last updated: 2017-12-04Bibliographically approved
Wu, C., Bai, L., Li, Z., Samuel, C. E., Akusjärvi, G. & Svensson, C. (2015). Poor growth of human adenovirus-12 compared to adenovirus-2 correlates with a failure to impair PKR activation during the late phase of infection. Virology, 475, 120-128
Open this publication in new window or tab >>Poor growth of human adenovirus-12 compared to adenovirus-2 correlates with a failure to impair PKR activation during the late phase of infection
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2015 (English)In: Virology, ISSN 0042-6822, E-ISSN 1096-0341, Vol. 475, p. 120-128Article in journal (Refereed) Published
Abstract [en]

Human adenovirus type 12 (HAdV-12) displays a relatively low virulence and slow replication in cultured human cells, which is manifested by premature death of HAdV-12-infected cells. Whereas HAdV-2 induction of IFN-beta expression was transient, HAdV-12-infected cells maintained high levels of IFN-beta expression, protein kinase R (PKR) activation and eIF-2 alpha phosphorylation throughout the infectious cycle. The importance of the IFN-inducible PKR kinase in restriction of HAdV-12 was supported by the enhanced growth of the virus following PKR knockdown in HeLa cells. Ectopic expression of HAdV-2 VA RNAI increased HAdV-12 hexon protein expression, suggesting that insufficient VA RNA expression contributes to the restricted growth of HAdV-12. Although some adenovirus species are known to persist in human lymphoid tissues, HAdV12 has so far not been found. Thus, it is possible that the inability of HAdV12 to evade the INF response may have implications for the virus to establish long-lasting or persistent infections. (C) 2014 Elsevier Inc. All rights reserved.

Keyword
Adenovirus, VA RNA, IFN, PKR, eIF2 alpha
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
urn:nbn:se:uu:diva-245366 (URN)10.1016/j.virol.2014.11.012 (DOI)000348176500012 ()25462352 (PubMedID)
Available from: 2015-02-26 Created: 2015-02-26 Last updated: 2017-12-04Bibliographically approved
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