Adenylyl cyclase mRNA localizes to the posterior of polarized DICTYOSTELIUM cells during chemotaxis

BMC Cell Biol. 2017 May 25;18(1):23. doi: 10.1186/s12860-017-0139-7.

Abstract

Background: In Dictyostelium discoideum, vesicular transport of the adenylyl cyclase A (ACA) to the posterior of polarized cells is essential to relay exogenous 3',5'-cyclic adenosine monophosphate (cAMP) signals during chemotaxis and for the collective migration of cells in head-to-tail arrangements called streams.

Results: Using fluorescence in situ hybridization (FISH), we discovered that the ACA mRNA is asymmetrically distributed at the posterior of polarized cells. Using both standard estimators and Monte Carlo simulation methods, we found that the ACA mRNA enrichment depends on the position of the cell within a stream, with the posterior localization of ACA mRNA being strongest for cells at the end of a stream. By monitoring the recovery of ACA-YFP after cycloheximide (CHX) treatment, we observed that ACA mRNA and newly synthesized ACA-YFP first emerge as fluorescent punctae that later accumulate to the posterior of cells. We also found that the ACA mRNA localization requires 3' ACA cis-acting elements.

Conclusions: Together, our findings suggest that the asymmetric distribution of ACA mRNA allows the local translation and accumulation of ACA protein at the posterior of cells. These data represent a novel functional role for localized translation in the relay of chemotactic signal during chemotaxis.

Keywords: Chemotaxis; Dictyostelium; Signal relay; mRNA.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenylyl Cyclases* / genetics
  • Adenylyl Cyclases* / metabolism
  • Animals
  • Cell Polarity / drug effects
  • Cell Polarity / genetics
  • Cells, Cultured
  • Chemotaxis / drug effects
  • Chemotaxis / genetics*
  • Cycloheximide / pharmacology
  • Cytoplasm / enzymology
  • Cytoplasmic Streaming / drug effects
  • Cytoplasmic Streaming / physiology
  • Dictyostelium / enzymology*
  • Dictyostelium / metabolism
  • In Situ Hybridization, Fluorescence
  • Protein Biosynthesis / drug effects
  • Protozoan Proteins* / genetics
  • Protozoan Proteins* / metabolism
  • RNA Transport / physiology
  • RNA, Messenger / analysis
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism*
  • RNA, Protozoan / analysis
  • RNA, Protozoan / genetics
  • RNA, Protozoan / metabolism
  • Regulatory Sequences, Ribonucleic Acid / physiology
  • Signal Transduction

Substances

  • Protozoan Proteins
  • RNA, Messenger
  • RNA, Protozoan
  • Regulatory Sequences, Ribonucleic Acid
  • Cycloheximide
  • Adenylyl Cyclases