Skip to main content Accessibility help
×
Home
Hostname: page-component-cf9d5c678-j7tnp Total loading time: 0.224 Render date: 2021-07-27T16:40:56.669Z Has data issue: true Feature Flags: { "shouldUseShareProductTool": true, "shouldUseHypothesis": true, "isUnsiloEnabled": true, "metricsAbstractViews": false, "figures": true, "newCiteModal": false, "newCitedByModal": true, "newEcommerce": true, "newUsageEvents": true }
RNA RNA

Article contents

Probing the structure of monomers and dimers of the bacterial virus phi29 hexamer RNA complex by chemical modification

Published online by Cambridge University Press:  01 September 2000

MARK TROTTIER
Affiliation:
Department of Pathobiology and Biochemistry and Molecular Biology Graduate Program, Purdue University, West Lafayette, Indiana 47907, USA Present address: ENH Research Institute, Evanston Hospital, Room B-620, 2650 Ridge Avenue, Evanston, Illinois 60201, USA.
YAHYA MAT-ARIP
Affiliation:
Department of Pathobiology and Biochemistry and Molecular Biology Graduate Program, Purdue University, West Lafayette, Indiana 47907, USA
CHUNLIN ZHANG
Affiliation:
Department of Pathobiology and Biochemistry and Molecular Biology Graduate Program, Purdue University, West Lafayette, Indiana 47907, USA Present address: Department of Virus Diseases, Division of CD & I, Walter Reed Army Institute of Research, Washington, DC 20307, USA.
CHAOPING CHEN
Affiliation:
Department of Pathobiology and Biochemistry and Molecular Biology Graduate Program, Purdue University, West Lafayette, Indiana 47907, USA Present address: Molecular Genetics and Biochemistry, University of Pittsburgh/Medical School, Pittsburgh, PA 15261, USA.
SITONG SHENG
Affiliation:
Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, Virginia 22908, USA
ZHIFENG SHAO
Affiliation:
Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, Virginia 22908, USA
PEIXUAN GUO
Affiliation:
Department of Pathobiology and Biochemistry and Molecular Biology Graduate Program, Purdue University, West Lafayette, Indiana 47907, USA
Get access

Abstract

All dsDNA viruses multiply their genome and assemble a procapsid, a protein shell devoid of DNA. The genome is subsequently inserted into the procapsid. The bacterial virus phi29 DNA translocating motor contains a hexameric RNA complex composed of six pRNAs. Recently, we found that pRNA dimers are building blocks of pRNA hexamers. Here, we report the structural probing of pRNA monomers and dimers by chemical modification under native conditions and in the presence or absence of Mg2+. The chemical-modification pattern of the monomer is compared to that of the dimer. The data strongly support the previous secondary-structure prediction of the pRNA concerning the single-stranded areas, including three loops and seven bulges. However, discrepancies between the modification patterns of two predicted helical regions suggest the presence of more complicated, higher-order structure in these areas. It was found that dimers were formed via hand-in-hand and head-to-head contact, as the interacting sequence of the right and left loops and all bases in the head loop were protected from chemical modification. Cryoatomic force microscopy revealed that the monomer displayed a check-mark shape and the dimer exhibited an elongated shape. The dimer was twice as long as the monomer. Direct observation of the shape and measurement of size and thickness of the images strongly support the conclusion from chemical modification concerning the head-to-head contact in dimer formation. Our results also suggest that the role for Mg2+ in pRNA folding is to generate a proper configuration for the right and head loops, which play key roles in this symmetrical head-to-head organization. This explains why Mg2+ plays a critical role in pRNA dimer formation, procapsid binding, and phi29 DNA packaging.

Type
Research Article
Information
RNA , Volume 6 , Issue 9 , September 2000 , pp. 1257 - 1266
Copyright
2000 RNA Society

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Send article to Kindle

To send this article to your Kindle, first ensure no-reply@cambridge.org is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about sending to your Kindle. Find out more about sending to your Kindle.

Note you can select to send to either the @free.kindle.com or @kindle.com variations. ‘@free.kindle.com’ emails are free but can only be sent to your device when it is connected to wi-fi. ‘@kindle.com’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

Find out more about the Kindle Personal Document Service.

Probing the structure of monomers and dimers of the bacterial virus phi29 hexamer RNA complex by chemical modification
Available formats
×

Send article to Dropbox

To send this article to your Dropbox account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your <service> account. Find out more about sending content to Dropbox.

Probing the structure of monomers and dimers of the bacterial virus phi29 hexamer RNA complex by chemical modification
Available formats
×

Send article to Google Drive

To send this article to your Google Drive account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your <service> account. Find out more about sending content to Google Drive.

Probing the structure of monomers and dimers of the bacterial virus phi29 hexamer RNA complex by chemical modification
Available formats
×
×

Reply to: Submit a response

Please enter your response.

Your details

Please enter a valid email address.

Conflicting interests

Do you have any conflicting interests? *