Mn2+ suppressor mutations and biochemical communication between Ty1 reverse transcriptase and RNase H domains

Robert M. Yarrington, Jichao Chen, Eric C. Bolton, Jef D. Boeke

Research output: Contribution to journalArticle

Abstract

Ty1 reverse transcriptase/RNase H (RT/RH) is exquisitely sensitive to manganese concentrations. Elevated intracellular free Mn2+ inhibits Ty1 retrotransposition and in vitro Ty1 RT-polymerizing activity. Furthermore, Mn2+ inhibition is not limited to the Ty1 RT, as this ion similarly inhibits the activities of both avian myeloblastosis virus and human immunodeficiency virus type 1 RTs. To further characterize Mn2+ inhibition, we generated RT/RH suppressor mutants capable of increased Ty1 transposition in pmr1Δ cells. PMR1 codes for a P-type ATPase that regulates intracellular calcium and manganese ion homeostasis, and pmr1 mutants accumulate elevated intracellular manganese levels and display 100-fold less transposition than PMR1+ cells. Mapping of these suppressor mutations revealed, surprisingly, that suppressor point mutations localize not to the RT itself but to the RH domain of the protein. Furthermore, Mn2+ inhibition of in vitro RT activity is greatly reduced in all the suppressor mutants, whereas RH activity and cleavage specificity remain largely unchanged. These intriguing results reveal that the effect of these suppressor mutations is transmitted to the polymerase domain and suggest biochemical communication between these two domains during reverse transcription.

Original languageEnglish (US)
Pages (from-to)9004-9012
Number of pages9
JournalJournal of Virology
Volume81
Issue number17
DOIs
StatePublished - Sep 2007

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Genetic Suppression
Ribonuclease H
RNA-directed DNA polymerase
RNA-Directed DNA Polymerase
Manganese
manganese
transposition (genetics)
mutants
Avian myeloblastosis virus
Avian Myeloblastosis Virus
Ions
ions
reverse transcription
point mutation
Human immunodeficiency virus 1
Point Mutation
polymerization
Reverse Transcription
adenosinetriphosphatase
Adenosine Triphosphatases

ASJC Scopus subject areas

  • Immunology

Cite this

Mn2+ suppressor mutations and biochemical communication between Ty1 reverse transcriptase and RNase H domains. / Yarrington, Robert M.; Chen, Jichao; Bolton, Eric C.; Boeke, Jef D.

In: Journal of Virology, Vol. 81, No. 17, 09.2007, p. 9004-9012.

Research output: Contribution to journalArticle

Yarrington, Robert M. ; Chen, Jichao ; Bolton, Eric C. ; Boeke, Jef D. / Mn2+ suppressor mutations and biochemical communication between Ty1 reverse transcriptase and RNase H domains. In: Journal of Virology. 2007 ; Vol. 81, No. 17. pp. 9004-9012.
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