The transmembrane domains of CAML are essential for a functional WRB/CAML receptor complex.

<p>(A) Schematic representation of CAML-Get2 chimeras. Position of transmembrane domains (TMD) are indicated. (B) <i>get1/get2</i> yeast cells carrying a genomically GFP-tagged version of Get3 were transformed with a plasmid containing the coding sequence of WRB in combination with CAML or CAML-Get2 chimeras. Subcellular Get3-GFP localization was analyzed by fluorescence microscopy. (C) <i>get1/get2</i> yeast cells were transformed with a plasmid containing the coding sequence of GFP-tagged Sed5 and Get1/Get2 or WRB in combination with CAML or CAML-Get2 chimeras. Subcellular GFP-Sed5 localization was analyzed by fluorescence microscopy. (D) Images taken in (C) were quantified to determine the distribution of fluorescence across bins of different pixel intensity for each strain. A minimum of 42 cells was analyzed per strain.</p

In combination, WRB and CAML rescue Get3 localization at the ER membrane and TA protein targeting.

<p>(A) <i>get1/get2</i> yeast cells carrying a genomically GFP-tagged version of Get3 were transformed with combinations of WRB, CAML, Get1 and Get2 encoding constructs. Subcellular Get3-GFP localization was analyzed by fluorescence microscopy. (B) <i>get1/get2</i> yeast cells were transformed with a plasmid containing the coding sequence of GFP-tagged Sed5 and combinations of WRB, CAML, Get1 and Get2 encoding constructs. Subcellular GFP-Sed5 localization was analyzed by fluorescence microscopy. (C) Images taken in (B) were quantified to determine the distribution of fluorescence across bins of different pixel intensity for each strain. A minimum of 41 cells was analyzed per strain.</p

Reflectometric Interference Spectroscopy (RIfS) and Surface plasmon resonance (SPR) measurements.

<p>Affinity constants of TRC40 and TRC40/RAMP4 complex for rough microsomes, WRBcc and CAMLcyt. Values represent the average calculated from three independent measurements.</p

Summary of the results obtained in this study.

<p>¹ growth rescue at 39°C.</p><p>‘n.a.’ = not assayed’.</p

WRB and CAML rescue the growth phenotypes of <i>get1/get2</i> yeast cells.

<p><i>get1/get2</i> yeast cells were transformed with combinations of WRB, CAML, Get1 and Get2 encoding constructs and serial dilutions spotted on different conditions: HC plates incubated at 30°C (control), 37°C+CuSO₄, 39°C, H₂O₂, hydroxyurea, tunicamycin, hygromycin.</p