Multiple regions of Kaposi's sarcoma-associated herpesvirus ORF59 RNA are required for its expression mediated by viral ORF57 and cellular RBM15

Maria Julia Massimelli; Vladimir Majerciak; Jeong-Gu Kang; David J Liewehr; Seth M Steinberg; Zhi-Ming Zheng

doi:10.3390/v7020496

Multiple regions of Kaposi's sarcoma-associated herpesvirus ORF59 RNA are required for its expression mediated by viral ORF57 and cellular RBM15

Viruses. 2015 Feb 3;7(2):496-510. doi: 10.3390/v7020496.

Authors

Maria Julia Massimelli¹, Vladimir Majerciak², Jeong-Gu Kang³, David J Liewehr⁴, Seth M Steinberg⁵, Zhi-Ming Zheng⁶

Affiliations

¹ Tumor Virus RNA Biology Section, Gene Regulation and Chromosome Biology Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 1050 Boyles Street, Frederick, MD 21702, USA. julia.massimelli@uci.edu.
² Tumor Virus RNA Biology Section, Gene Regulation and Chromosome Biology Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 1050 Boyles Street, Frederick, MD 21702, USA. majerciv@mail.nih.gov.
³ Tumor Virus RNA Biology Section, Gene Regulation and Chromosome Biology Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 1050 Boyles Street, Frederick, MD 21702, USA. jkang@jkbiopharma.com.
⁴ Biostatistics & Data Management Section, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. liewehrd@mail.nih.gov.
⁵ Biostatistics & Data Management Section, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. steinbes@mail.nih.gov.
⁶ Tumor Virus RNA Biology Section, Gene Regulation and Chromosome Biology Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 1050 Boyles Street, Frederick, MD 21702, USA. zhengt@exchange.nih.gov.

Abstract

KSHV ORF57 (MTA) promotes RNA stability of ORF59, a viral DNA polymerase processivity factor. Here, we show that the integrity of both ORF59 RNA ends is necessary for ORF57-mediated ORF59 expression and deletion of both 5' and 3' regions, or one end region with a central region, of ORF59 RNA prevents ORF57-mediated translation of ORF59. The ORF59 sequence between nt 96633 and 96559 resembles other known MTA-responsive elements (MREs). ORF57 specifically binds to a stem-loop region from nt 96596-96572 of the MRE, which also binds cellular RBM15. Internal deletion of the MRE from ORF59 led to poor export, but accumulation of nuclear ORF59 RNA in the presence of ORF57 or RBM15. Despite of being translatable in the presence of ORF57, this deletion mutant exhibits translational defect in the presence of RBM15. Together, our results provide novel insight into the roles of ORF57 and RBM15 in ORF59 RNA accumulation and protein translation.

Publication types

Research Support, N.I.H., Intramural

MeSH terms

Base Sequence
Cell Line
Cells, Cultured
Epistasis, Genetic
Gene Expression Regulation, Viral*
Herpesviridae Infections / metabolism
Herpesviridae Infections / virology
Herpesvirus 8, Human / genetics*
Humans
Molecular Sequence Data
Nucleic Acid Conformation
Protein Biosynthesis
RNA Transport
RNA, Viral / chemistry
RNA, Viral / genetics*
RNA, Viral / metabolism
RNA-Binding Proteins / metabolism*
Response Elements
Viral Proteins / chemistry
Viral Proteins / genetics*
Viral Regulatory and Accessory Proteins / chemistry
Viral Regulatory and Accessory Proteins / genetics
Viral Regulatory and Accessory Proteins / metabolism*

Substances

ORF57 protein, human herpesvirus 8
ORF59 protein, Human herpesvirus 8
RBM15 protein, human
RNA, Viral
RNA-Binding Proteins
Viral Proteins
Viral Regulatory and Accessory Proteins

Grants and funding

Intramural NIH HHS/United States