Summary.
The early region 4 (E4) of porcine adenovirus 3 (PAdV-3) was characterized by Northern blot, rapid amplification of cDNA ends (RACE), RT-PCR and cDNA sequence analysis. Northern blot analysis revealed three different classes of transcripts, which appeared and peaked at different times post-infection. The RT-PCR, RACE and cDNA sequence analysis identified nine major E4 transcripts, all of which shared a 107-bp 5′ leader sequence and a 126-bp 3′ terminus. These transcripts have one to three introns removed. Interestingly, of the nine major transcripts, there was one fusion transcript of ORFp1 and ORFp7 (ORFp1/7), which codes for a protein of 119 amino acids. All transcripts initiated at nucleotide 33740 of the PAdV-3 genome. To identify proteins, rabbit antiserum was prepared using a bacterial fusion protein encoding p2, p3, p4 or p7 proteins. Serum against p2, p3 and p4 immunoprecipitated proteins of 13.5, 13.6 and 15.3 kDa, respectively, in in-vitro transcribed and translated mRNA and in PAdV-3-infected cells. Serum against p7 immunoprecipitated a protein of 19.8 kDa in in-vitro transcription and translation analysis but recognized two proteins of 19.8 kDa (encoded by ORFp7) and 14 kDa (encoded by the fusion transcript ORF1/7) in PAdV-3-infected cells. The protein encoded by ORFp2 was localized in the nucleus of PAdV-3-infected cells. The proteins encoded by ORFp3 and ORFp7\ORFp1/7 were detected in the cytoplasm of PAdV-3-infected cells. However, the protein encoded by ORFp4 was observed both in the cytoplasm and nucleus of PAdV-3-infected cells.
Similar content being viewed by others
References
D Armentano J Zabner C Sacks CC Sookdeo MP Smith JA St George SC Wadsworth AE Smith RJ Gregory (1997) ArticleTitleEffect of the E4 region on the persistence of transgene expression from adenovirus vectors J Virol 71 2408–2416 Occurrence Handle9032378 Occurrence Handle1:CAS:528:DyaK2sXht1Wrtrc%3D
MK Baxi LA Babiuk M Mehtali SK Tikoo (1999) ArticleTitleTranscription map and expression of bovine herpesvirus-1 glycoprotein D in early region 4 of bovine adenovirus-3 Virology 261 143–152 Occurrence Handle10441562 Occurrence Handle10.1006/viro.1999.9826 Occurrence Handle1:CAS:528:DyaK1MXltVagtLw%3D
GW Both (2004) ArticleTitleOvine atadenovirus: a review of its biology, biosafety profile and application as a gene delivery vector Immunol Cell Biol 82 189–195 Occurrence Handle15061773 Occurrence Handle10.1046/j.0818-9641.2004.01223.x Occurrence Handle1:CAS:528:DC%2BD2cXksFOksbo%3D
LT Chow TR Broker JB Lewis (1979) ArticleTitleComplex splicing patterns of RNAs from the early regions of adenovirus-2 J Mol Biol 134 265–303 Occurrence Handle537065 Occurrence Handle10.1016/0022-2836(79)90036-6 Occurrence Handle1:CAS:528:DyaL3cXjs1aiuw%3D%3D
JR Cutt T Shenk P Hearing (1987) ArticleTitleAnalysis of adenovirus early region 4-encoded polypeptides synthesized in productively infected cells J Virol 61 543–552 Occurrence Handle2949089 Occurrence Handle1:CAS:528:DyaL2sXhtFKiu7w%3D
JB Derbyshire MC Clarke AP Collins (1975) ArticleTitleSerological and pathogenicity studies with some unclassified porcine adenoviruses J Comp Pathol 85 437–443 Occurrence Handle167059 Occurrence Handle10.1016/0021-9975(75)90031-6 Occurrence Handle1:STN:280:CSqC1cbptl0%3D
I Dix KN Leppard (1995) ArticleTitleExpression of adenovirus type 5 E4 Orf2 protein during lytic infection J Gen Virol 76 1051–1055 Occurrence Handle9049357 Occurrence Handle1:CAS:528:DyaK2MXksl2ns7o%3D Occurrence Handle10.1099/0022-1317-76-4-1051
B Falgout G Ketner (1987) ArticleTitleAdenovirus early region 4 is required for efficient virus particle assembly J Virol 61 3759–3768 Occurrence Handle2824814 Occurrence Handle1:CAS:528:DyaL1cXhvFSgsw%3D%3D
Francki RB, Fauquet CM, Knudson DL, Brown F (1991) Classification and nomenclature of viruses. Fifth Report of the International Committee on Taxonomy of Viruses. In: Arch Virol Suppl 2, Springer Wien, New York, pp 140–144
P Gilardi M Perricaudet (1986) ArticleTitleThe E4 promoter of adenovirus type 2 contains an E1A dependent cis-acting element Nucleic Acids Res 14 9035–9049 Occurrence Handle2947044 Occurrence Handle10.1093/nar/14.22.9035 Occurrence Handle1:CAS:528:DyaL2sXivVemtg%3D%3D
FD Goodrum T Shenk DA Ornelles (1996) ArticleTitleAdenovirus early region 4 34-kilodalton protein directs the nuclear localization of the early region 1B 55-kilodalton protein in primate cells J Virol 70 6323–6335 Occurrence Handle8709260 Occurrence Handle1:CAS:528:DyaK28XkvFGgsbo%3D
DA Haig MC Clarke MS Pereira (1964) ArticleTitleIsolation of an adenovirus from a pig J Comparat Pathol 74 81–84 Occurrence Handle1:STN:280:CCuD283pt1w%3D
T Hirahara H Yasuhara O Matsui S Fukuyama M Yamanaka A Izumida K Yoshiki K Kodama M Nakai N Sasaki (1990) ArticleTitleGrowth activity of porcine adenoviruses in primary porcine testicular cell cultures Nippon Juigaku Zasshi 52 1089–1091 Occurrence Handle2280492 Occurrence Handle1:STN:280:By6C38vmtlI%3D
JM Kaplan D Armentano A Scaria LA Woodworth SE Pennington SC Wadsworth AE Smith RJ Gregory (1999) ArticleTitleNovel role for E4 region genes in protection of adenovirus vectors from lysis by cytotoxic T lymphocytes J Virol 73 4489–4492 Occurrence Handle10196353 Occurrence Handle1:CAS:528:DyaK1MXis1eqsr0%3D
A Khatri GW Both (1998) ArticleTitleIdentification of transcripts and promoter regions of ovine adenovirus OAV287 Virology 245 128–141 Occurrence Handle9614874 Occurrence Handle10.1006/viro.1998.9136 Occurrence Handle1:CAS:528:DyaK1cXjsFWgu70%3D
SC Kring AO Ball KR Spindler (1992) ArticleTitleTranscription mapping of mouse adenovirus type 1 early region 4 Virology 190 248–255 Occurrence Handle1388309 Occurrence Handle10.1016/0042-6822(92)91211-C Occurrence Handle1:CAS:528:DyaK38XlvVGqsb8%3D
V Kulshrestha LA Babiuk SK Tikoo (2004) ArticleTitleRole of bovine adenovirus-3 33 K protein in viral replication Virology 323 59–69 Occurrence Handle10.1016/j.virol.2004.02.024 Occurrence Handle1:CAS:528:DC%2BD2cXksVemtL0%3D
JB Lee MK Baxi N Idamakanti PS Reddy AN Zakhartchouk C Pyne LA Babiuk SK Tikoo (1998) ArticleTitleGenetic organization and DNA sequence of early region 4 of bovine adenovirus type 3 Virus Genes 17 99–100 Occurrence Handle9778793 Occurrence Handle10.1023/A:1008017404513 Occurrence Handle1:CAS:528:DyaK1cXmt1yhtbw%3D
KN Leppard (1997) ArticleTitleE4 gene function in adenovirus, adenovirus vector and adeno-associated virus infections J Gen Virol 78 2131–2138 Occurrence Handle9291998 Occurrence Handle1:CAS:528:DyaK2sXls1Oitr8%3D
X Li LA Babiuk SK Tikoo (2004) ArticleTitleAnalysis of early region 4 of porcine adenovirus type 3 Virus Res 104 181–190 Occurrence Handle15246655 Occurrence Handle10.1016/j.virusres.2004.04.011 Occurrence Handle1:CAS:528:DC%2BD2cXls1ajtro%3D
T Matsuo WS Wold S Hashimoto A Rankin J Symington M Green (1982) ArticleTitlePolypeptides encoded by transforming region E 1b of human adenovirus 2: immunoprecipitation from transformed and infected cells and cell-free translation of E 1b-specific mRNA Virology 118 456–465 Occurrence Handle7090186 Occurrence Handle10.1016/0042-6822(82)90366-X Occurrence Handle1:CAS:528:DyaL38XhvFChurk%3D
M Moore N Horikoshi T Shenk (1996) ArticleTitleOncogenic potential of the adenovirus E4orf6 protein Proc Natl Acad Sci USA 93 11295–11301 Occurrence Handle8876129 Occurrence Handle10.1073/pnas.93.21.11295 Occurrence Handle1:CAS:528:DyaK28Xmtlahurg%3D
JR Nevins HS Ginsberg JM Blanchard MC Wilson JE Darnell SuffixJr (1979) ArticleTitleRegulation of the primary expression of the early adenovirus transcription units J Virol 32 727–733 Occurrence Handle513202 Occurrence Handle1:CAS:528:DyaL3cXnt1CgsQ%3D%3D
U Pettersson (1986) ArticleTitleAdenovirus gene expression and replication: a history review Cancer Cells 4 37–57 Occurrence Handle1:CAS:528:DyaL2sXhtFCgsr8%3D
PS Reddy N Idamakanti JB Derbyshire E Nagy (1997) ArticleTitleCharacterization of the early region 4 of porcine adenovirus type 3 Virus Genes 15 87–90 Occurrence Handle9354276 Occurrence Handle10.1023/A:1007935720501 Occurrence Handle1:CAS:528:DyaK2sXmslKitb0%3D
PS Reddy N Idamakanti BH Hyun SK Tikoo LA Babiuk (1999) ArticleTitleDevelopment of porcine adenovirus-3 as an expression vector J Gen Virol 80 563–570 Occurrence Handle10091994 Occurrence Handle1:CAS:528:DyaK1MXhvVymt7w%3D
PS Reddy N Idamakanti JY Song JB Lee BH Hyun JH Park SH Cha YT Bae SK Tikoo LA Babiuk (1998) ArticleTitleNucleotide sequence and transcription map of porcine adenovirus type 3 Virology 251 414–426 Occurrence Handle9837805 Occurrence Handle10.1006/viro.1998.9418 Occurrence Handle1:CAS:528:DyaK1cXnvVGjurc%3D
Regenmortel V (2000) Virus taxonomy. The Seventh Report of the International Committee on Taxonomy of Viruses. Academic Press, San Diego
D Ross E Ziff (1992) ArticleTitleDefective synthesis of early region 4 mRNAs during abortive adenovirus infections in monkey cells J Virol 66 3110–3117 Occurrence Handle1313921 Occurrence Handle1:CAS:528:DyaK38XisFajsLo%3D
DT Rowe FL Graham PE Branton (1983) ArticleTitleIntracellular localization of adenovirus type 5 tumor antigens in productively infected cells Virology 129 456–468 Occurrence Handle6353747 Occurrence Handle10.1016/0042-6822(83)90183-6 Occurrence Handle1:CAS:528:DyaL3sXlslCjtb8%3D
F Sanger S Nicklen AR Coulson (1977) ArticleTitleDNA sequencing with chain-terminating inhibitors Proc Natl Acad Sci USA 74 5463–5467 Occurrence Handle271968 Occurrence Handle10.1073/pnas.74.12.5463 Occurrence Handle1:CAS:528:DyaE1cXhtlaru7Y%3D
P Sarnow P Hearing CW Anderson DN Halbert T Shenk AJ Levine (1984) ArticleTitleAdenovirus early region 1B 58,000-dalton tumor antigen is physically associated with an early region 4 25,000-dalton protein in productively infected cells J Virol 49 692–700 Occurrence Handle6699935 Occurrence Handle1:CAS:528:DyaL2cXhtlGmsro%3D
T Shenk (1996) Adenoviridae: The viruses and their replication BN Fields DM Knipe PM Howley (Eds) et al. Fields virology Lippincott-Raven Philadephia
B Tauber T Dobner (2001) ArticleTitleMolecular regulation and biological function of adenovirus early genes: the E4 ORFs Gene 278 1–23 Occurrence Handle11707318 Occurrence Handle10.1016/S0378-1119(01)00722-3 Occurrence Handle1:CAS:528:DC%2BD3MXot1KnsLs%3D
MA Tigges HJ Raskas (1984) ArticleTitleSplice junctions in adenovirus 2 early region 4 mRNAs: multiple splice sites produce 18 to 24 RNAs J Virol 50 106–117 Occurrence Handle6336328 Occurrence Handle1:CAS:528:DyaL2cXhvVKmtrY%3D
T Tuboly M Nagy E Nagy (2002) ArticleTitleCharacterization of early region 4 of porcine adenovirus serotype 5 Virus Genes 20 217–219 Occurrence Handle10.1023/A:1008136411020
A Virtanen P Gilardi A Naslund JM LeMoullec U Pettersson M Perricaudet (1984) ArticleTitlemRNAs from human adenovirus 2 early region 4 J Virol 51 822–831 Occurrence Handle6088804 Occurrence Handle1:CAS:528:DyaL2cXlvVGjsb8%3D
H Watanabe T Imai PA Sharp H Handa (1988) ArticleTitleIdentification of two transcription factors that bind to specific elements in the promoter of the adenovirus early-region 4 Mol Cell Biol 8 1290–1300 Occurrence Handle3367909 Occurrence Handle1:CAS:528:DyaL1cXhsVCgtbs%3D
S Weigel M Dobbelstein (2000) ArticleTitleThe nuclear export signal within the E4orf6 protein of adenovirus type 5 supports virus replication and cytoplasmic accumulation of viral mRNA J Virol 74 764–772 Occurrence Handle10623738 Occurrence Handle10.1128/JVI.74.2.764-772.2000 Occurrence Handle1:CAS:528:DC%2BD3cXkt1ClsA%3D%3D
DH Weinberg G Ketner (1986) ArticleTitleAdenoviral early region 4 is required for efficient viral DNA replication and for late gene expression J Virol 57 833–838 Occurrence Handle3485200 Occurrence Handle1:CAS:528:DyaL28Xht1CjsLs%3D
RS Weiss SS Lee BV Prasad RT Javier (1997) ArticleTitleHuman adenovirus early region 4 open reading frame 1 genes encode growth-transforming proteins that may be distantly related to dUTP pyrophosphatase enzymes J Virol 71 1857–1870 Occurrence Handle9032316 Occurrence Handle1:CAS:528:DyaK2sXht1Wmtbo%3D
Y Zhou C Pyne SK Tikoo (2001) ArticleTitleCharacterization of DNA binding protein of porcine adenovirus type 3 Intervirology 44 350–354 Occurrence Handle11805441 Occurrence Handle10.1159/000050070 Occurrence Handle1:CAS:528:DC%2BD38XmtFOjug%3D%3D
Y Zhou SK Tikoo (2001) ArticleTitleAnalysis of early region 1 of porcine adenovirus type 3 Virology 291 68–76 Occurrence Handle11878877 Occurrence Handle10.1006/viro.2001.1219 Occurrence Handle1:CAS:528:DC%2BD3MXovFGgsrk%3D
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Li, X., Babiuk, L. & Tikoo, S. Transcription mapping and characterization of proteins produced from early region 4 of porcine adenovirus type 3. Arch Virol 152, 495–505 (2007). https://doi.org/10.1007/s00705-006-0876-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00705-006-0876-y