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[Translational frameshift may be occur in p11, an interaction protein of Cx31, in yeast]

Liang-Qun Huang1, Xiao-Liu Shi, Zhi-Ping Tan

  • 1National Laboratory of Medical Genetics of China, Changsha 410078, China. nlmglcy@public.cs.hn.cn

Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao Acta Biochimica Et Biophysica Sinica
|May 15, 2002
PubMed

Insights

Connexin 31 interacts with the p11 protein, a subunit of annexin II. This interaction is mediated by a specific domain on connexin 31 and involves a translational frameshift in p11 expression, as identified through yeast two-hybrid screening.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Protein Interactions

Background:

  • Connexin 31 (Cx31) is a gap junction protein with a carboxy-terminal domain containing phosphorylation sites.
  • The p11 protein is a member of the S100 family and a subunit of the annexin II tetramer.

Purpose of the Study:

  • To identify proteins that interact with the carboxy-terminus of connexin 31.
  • To characterize the binding domain and mechanism of interaction between Cx31 and identified proteins.

Main Methods:

  • Yeast two-hybrid protein interaction screen.
  • Construction of expression plasmids for connexin 31 C-terminus and p11.
  • In vitro binding assays using purified recombinant proteins.

Main Results:

  • The p11 protein was identified as a binding partner for connexin 31.
  • A translational frameshift in p11 expression was observed, with the open reading frame (ORF) binding Cx31, but not the 5' UTR.
  • The p11 binding domain on connexin 31 was localized to codons 206-237.
  • In vitro assays confirmed direct interaction between recombinant Cx31 and p11.

Conclusions:

  • Connexin 31 directly interacts with the p11 protein.
  • The interaction is dependent on the p11 open reading frame and a specific region of the connexin 31 C-terminus.
  • A translational frameshift mechanism influences the interaction between Cx31 and p11.

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