Table 4. Transition zone and IFT components required for trafficking and localization of GPCR::GFP fusion proteins to AWB and ASK cilia.
Strainb | Fusion protein | Neuron | % localized to:a | ||||||
---|---|---|---|---|---|---|---|---|---|
Cilium | Cilium, PCMC/distal dendrite | PCMC/Distal dendritec | Cilium, dendrite, cell body | Cell body | No expression | P-valued | |||
Wild type | STR-44 | AWB | 100 | 0 | 0 | 0 | 0 | 0 | |
STR-163 | AWB | 100 | 0 | 0 | 0 | 0 | 0 | ||
STR-163 | ASK | 68 | 0 | 0 | 0 | 11 | 21 | ||
SRBC-64 | ASK | 0 | 95 | 0 | 0 | 0 | 5 | ||
TZ + inversin compartment | |||||||||
mks-1(tm2705); mksr-2(tm2452); mksr-1(ok2092) | STR-44 | AWB | 100 | 0 | 0 | 0 | 0 | 0 | |
STR-163 | AWB | 100 | 0 | 0 | 0 | 0 | 0 | ||
STR-163 | ASK | 48 | 0 | 0 | 0 | 19 | 33 | ||
nphp-4(tm925) | STR-44 | AWB | 100 | 0 | 0 | 0 | 0 | 0 | |
STR-163 | AWB | 100 | 0 | 0 | 0 | 0 | 0 | ||
STR-163 | ASK | 85 | 0 | 0 | 0 | 0 | 15 | ||
mks-5(tm3100) | STR-44e | AWB | 0 | 100 | 0 | 0 | 0 | 0 | <0.001 |
STR-163e | AWB | 0 | 100 | 0 | 0 | 0 | 0 | <0.001 | |
STR-163 | ASK | 52 | 0 | 0 | 0 | 15 | 33 | ||
Ex[str-1p::mks-5] | STR-44 | AWB | 65e | 35 | 0 | 0 | 0 | 0 | <0.001f |
nphp-2(gk653) | STR-44 | AWB | 100 | 0 | 0 | 0 | 0 | 0 | |
STR-163 | AWB | 100 | 0 | 0 | 0 | 0 | 0 | ||
STR-163 | ASK | 68 | 0 | 0 | 0 | 9 | 23 | ||
SRBC-64 | ASK | 0 | 95 | 0 | 0 | 0 | 5 | ||
IFT related | |||||||||
osm-3(p802) | STR-44 | AWB | 100 | 0 | 0 | 0 | 0 | 0 | |
STR-163 | AWB | 100 | 0 | 0 | 0 | 0 | 0 | ||
STR-163 | ASK | 72 | 0 | 0 | 0 | 14 | 14 | ||
kap-1(ok676) | STR-44 | AWB | 100 | 0 | 0 | 0 | 0 | 0 | |
STR-163 | AWB | 100 | 0 | 0 | 0 | 0 | 0 | ||
STR-163 | ASK | 67 | 0 | 0 | 0 | 14 | 19 | ||
tub-1(nr2004) | STR-44e | AWB | 0 | 100 | 0 | 0 | 0 | 0 | <0.001 |
STR-163e | AWB | 0 | 100 | 0 | 0 | 0 | 0 | <0.001 | |
STR-163e | ASK | 0 | 0 | 81 | 0 | 0 | 19 | <0.001 | |
SRBC-64 | ASK | 0 | 100 | 0 | 0 | 0 | 0 | ||
tub-1(nr2044) | STR-163e | AWB | 0 | 100 | 0 | 0 | 0 | 0 | <0.001 |
daf-10(e1387) | STR-44e | AWB | 0 | 100 | 0 | 0 | 0 | 0 | <0.001 |
STR-163e | AWB | 0 | 100 | 0 | 0 | 0 | 0 | <0.001 | |
STR-163 | ASK | 62 | 0 | 0 | 0 | 24 | 14 | ||
SRBC-64 | ASK | 0 | 0 | 100 | 0 | 0 | 0 | <0.001 | |
daf-10(p821) | STR-44e | AWB | 0 | 100 | 0 | 0 | 0 | 0 | <0.001 |
SRBC-64 | ASK | 0 | 0 | 100 | 0 | 0 | 0 | <0.001 |
Adult animals grown at 20° were examined. n ≥ 20 animals each. Expression from the same array was examined in wild-type and mutant strains except for the mks-1; mksr-2; mksr-1 triple mutant strain, which was injected with the indicated fusion constructs. TZ, transition zone.
Localization patterns were examined in animals exhibiting relatively wild-type ciliary morphology with the exception of mks-5, tub-1, daf-10, and osm-3 mutants. AWB cilia are truncated in mks-5 and tub-1 mutants; AWB and ASK cilia are truncated in daf-10 mutants; ASK cilia are truncated in osm-3 mutants.
Mutant strains were examined together with wild type in each experiment.
Weak expression was observed in cilia.
Differences among proportions in different categories were compared with respective wild-type values unless indicated otherwise in the relevant neuron type for statistical significance. P-values were determined using a χ2 test of independence. Only differences at P < 0.05 or lower are indicated.
Cilia in these strains were visualized via expression of str-1p::mCherry (AWB) or srbc-66p::che-13::TagRFP (ASK).
As compared to values in mks-5 mutants. The AWB ciliary morphological defects were also rescued (see Figure 4B).