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Pat1 contributes to the RNA binding activity of the Lsm1-7-Pat1 complex

Ashis Chowdhury1, Swathi Kalurupalle1, Sundaresan Tharun2

  • 1Department of Biochemistry, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814, USA.

RNA (New York, N.Y.)
|July 19, 2014
PubMed

Insights

The Lsm1-7-Pat1 complex is essential for mRNA decay in eukaryotes. Pat1 protein cooperates with the Lsm1-7 complex to bind RNA, enabling efficient mRNA degradation.

Area of Science:

  • Molecular Biology
  • RNA Biology
  • Biochemistry

Background:

  • mRNA decay is a crucial process in eukaryotes, with decapping being a rate-limiting step.
  • The hetero-octameric Lsm1-7-Pat1 complex is vital for efficient mRNA decapping in vivo.
  • This complex exhibits a specific preference for binding oligoadenylated RNAs, which is critical for its function.

Purpose of the Study:

  • To elucidate the specific role of the Pat1 protein in the RNA binding and functional capabilities of the Lsm1-7-Pat1 complex.
  • To investigate the contribution of Pat1 to the complex's unique preference for oligoadenylated RNAs.
  • To determine the structural basis for Pat1-Lsm1-7 interaction and its impact on complex function.

Main Methods:

  • Comparative analysis of RNA binding properties of wild-type Lsm1-7-Pat1 complex, Lsm1-7 complex alone, and purified Pat1 fragments.
  • In vitro reconstitution of the Lsm1-7-Pat1 complex from its components.
  • Investigation of direct RNA-Pat1 interactions within the reconstituted complex.
  • Assessment of Pat1-Lsm1-7 interaction in vivo, focusing on the role of Pat1 domains.

Main Results:

  • The Lsm1-7 complex and Pat1 fragments alone exhibit significantly reduced RNA binding activity and impaired recognition of oligo(A) tails.
  • Reconstitution of the Lsm1-7-Pat1 complex restores robust RNA binding and oligo(A) tail recognition.
  • Pat1 directly contacts RNA when part of the Lsm1-7-Pat1 complex.
  • The middle domain of Pat1 is essential for its in vivo interaction with the Lsm1-7 complex.

Conclusions:

  • The mRNA decay function and unique RNA binding properties of the Lsm1-7-Pat1 complex arise from the cooperative interaction between Pat1 and the Lsm1-7 protein ring.
  • Pat1 plays a direct and essential role in the complex's ability to bind and recognize specific RNA structures.
  • Structural integrity and cooperation between Pat1 and Lsm1-7 are critical for efficient mRNA decapping and subsequent degradation.
Keywords:
Lsm1-7–Pat1 complexPat1Sm-like proteinsdecappingmRNA decay

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